CLIAMS:1. An automated bacterial challenge test skid for preventing test invalidation owing to failure of a single filter, the automated bacterial challenge test skid comprising:
a first automated process control loop, the first automated process control loop comprising a first vessel, the first vessel comprising an inlet port and an outlet port, the inlet port coupled to a peristaltic pump to circulate a solution through a first sterilizing grade test filter, the first sterilizing grade test filter coupled to a first recovery filter, and the outlet port operable to receive filtered contents from the first recovery filter;
a second automated process control loop, the second automated process control loop comprising a second vessel, the second vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a second sterilizing grade test filter, the second sterilizing grade test filter coupled to a second recovery filter, and the outlet port operable to receive filtered contents from the second recovery filter;
a third automated process control loop, the third automated process control loop comprising a third vessel, the third vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a third sterilizing grade test filter, the third sterilizing grade test filter coupled to a third recovery filter, and the outlet port operable to receive filtered contents from the third recovery filter; and
a fourth automated process control loop, the fourth automated process control loop comprising a fourth vessel, the fourth vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a fourth sterilizing grade test filter, the fourth sterilizing grade test filter coupled to a fourth recovery filter, and the outlet port operable to receive filtered contents from the fourth recovery filter, wherein the first automated process control loop in conjunction with the second automated process control loop, the third automated process control loop, and the fourth automated process control loop prevents test invalidation owing to failure of a single filter.

2. The automated bacterial challenge test skid of claim 1,wherein the first automated process control loop in conjunction with the second automated process control loop, the third automated process control loop, and the fourth automated process control loop prevents test invalidation owing to contamination of a single vessel.

3. The automated bacterial challenge test skid of claim 1, further comprising a cryo-bath for maintaining an even distribution of temperature across one or more vessels.

4. The automated bacterial challenge test skid of claim 1, wherein the first automated process control loop, the second automated process control loop, the third automated process control loop, and the fourth automated process control loop enable automatic control of one or more process parameters, wherein the one or more process parameters comprises at least one of temperature, pressure, differential pressure, contact time, flow rate, and total volume filtered.

5. The automated bacterial challenge test skid of claim 4, wherein a change in the one or more process parameters are captured to provide a 21 CFR part 11 compliant audit trail.

6. An automated bacterial challenge test skid for open source validation and operable with a plurality of filter-make, the automated bacterial challenge test skid comprising:

a first automated process control loop, the first automated process control loop comprising a first vessel, the first vessel comprising an inlet port and an outlet port, the inlet port coupled to a peristaltic pump to circulate a solution through a first sterilizing grade test filter, the first sterilizing grade test filter coupled to a first recovery filter, and the outlet port operable to receive filtered contents from the first recovery filter;
a second automated process control loop, the second automated process control loop comprising a second vessel, the second vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a second sterilizing grade test filter, the second sterilizing grade test filter coupled to a second recovery filter, and the outlet port operable to receive filtered contents from the second recovery filter;
a third automated process control loop, the third automated process control loop comprising a third vessel, the third vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a third sterilizing grade test filter, the third sterilizing grade test filter coupled to a third recovery filter, and the outlet port operable to receive filtered contents from the third recovery filter; and
a fourth automated process control loop, the fourth automated process control loop comprising a fourth vessel, the fourth vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a fourth sterilizing grade test filter, the fourth sterilizing grade test filter coupled to a fourth recovery filter, and the outlet port operable to receive filtered contents from the fourth recovery filter, wherein the first automated process control loop in conjunction with the second automated process control loop, the third automated process control loop, and the fourth automated process control loop allows validation of one or more sterilizing filter-make provided by a customer for bacterial retention capability.

7. The automated bacterial challenge test skid of claim 6, wherein the first automated process control loop in conjunction with the second automated process control loop, the third automated process control loop, and the fourth automated process control loop prevents test invalidation owing to one or more of the following conditions:
if one or more of vessels are contaminated, and
if integrity of one or more filters is compromised.

8. The automated bacterial challenge test skid of claim 6, further comprising a cryo-bath for maintaining an even distribution of temperature across one or more vessels.

9. The automated bacterial challenge test skid of claim 6, wherein the first automated process control loop, the second automated process control loop, the third automated process control loop, and the fourth automated process control loop enable automatic control of one or more process parameters, wherein the one or more process parameters comprises at least one of temperature, pressure, differential pressure, contact time, flow rate, and total volume filtered.

10. The automated bacterial challenge test skid of claim 9, wherein a change in the one or more process parameters are captured to provide a 21 CFR part 11 compliant audit trail.
,TagSPECI:
FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
As amended by the Patents (Amendment) Act, 2005
&
The Patents Rules, 2003
As amended by the Patents (Amendment) Rules, 2006
COMPLETE SPECIFICATION
(See section 10 and rule 13)

TITLE OF THE INVENTION
MULTI-TEST AUTOMATED BACTERIAL CHALLENGE SKID
APPLICANT
Bangalore Biotech Labs Pvt. ltd.
49/2, Gubbi Cross, Hennur Bagalur Road, Kothanur P.O., Bangalore 560 077, INDIA

INVENTORS Vibin Baby Joseph, an Indian national, of Bangalore Biotech Labs Pvt. ltd, 49/2, Gubbi Cross, Hennur Bagalur Road, Kothanur P.O, Bangalore 560 077
Maqsood Ali, an Indian national of Bangalore Biotech Labs Pvt. ltd, 49/2, Gubbi Cross, Hennur Bagalur Road, Kothanur P.O, Bangalore 560 077, and
Subramanian Salavadi Easwaran, an Indian national, of Bangalore Biotech Labs Pvt. ltd, 49/2, Gubbi Cross, Hennur Bagalur Road, Kothanur P.O, Bangalore 560 077

PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the nature of this invention and the manner in which it is to be performed.

MULTI-TEST AUTOMATED BACTERIAL CHALLENGE SKID

FIELD OF THE INVENTION

[1] The present invention relates to the field of filter validation studies, and more specifically to bacterial challenge test for validating a filter’s retention capabilities.

BACKGROUND

[2] Bacterial challenge test is used to validate a membrane filter and prove its retention capabilities. Validation is performed to test whether the membrane filter is able to retain the challenge organism, Brevundimonas diminuta (American Type Culture Collection - ATCC 19146), and give a sterile filtrate. Typically, Brevundimonas diminuta (ATCC 19146) is used as the standard micro-organism for validation of sterilizing-grade membrane filters. During bacterial challenge test, the solution is inoculated with Brevundimonas diminuta (ATCC 19146) at a concentration =1x107 CFU /cm2 of effective filtration area. This test is performed at the conditions specified by the Biotech and Pharmaceutical customers, thereby making the test an actual simulation of the processes that are followed at the shop-floor during filtration/aseptic filling of a given drug product. The conditions that are to be monitored are the process temperature, pressure, differential pressure, flow rate, total contact time and the total volume filtered. Further, Bacterial challenge test involves establishing the viability of the test organism in the given test solution.

[3] Existing techniques for bacterial challenge test involves a test jig with a single vessel for containing the bacteria inoculated product solution and passing the solution through one or more test filters by making use of a pump and a plurality of control valves. Using a single vessel for low volumes of the solution will bring about uneven temperature distribution across the solution. Further, the cleaning time and solution required will be high. Moreover, if there is contamination in the vessel, the entire test becomes invalid. Since the source of the solution is same, i.e. as it is coming from a single vessel, the entire test becomes invalid if the integrity of the filters is compromised. Another concern of the existing bacterial challenge tests is that the process parameters are controlled through manual operation. Often, the user desires to have an automated control of the process parameters which is compliant with 21 CFR part 11 guidelines for audit trail.

[4] In light of the foregoing discussion, there is a need for an automated system for performing bacterial challenge test at the same time providing a fail-proof system to test the retention capability of the filter membrane.

SUMMARY

[5] The above-mentioned needs are met by an automated bacterial challenge test skid that provides a fail-proof system to test the bacterial retention capability of the filter membrane.

[6] An example of an automated bacterial challenge test skid for preventing test invalidation owing to failure of a single filter includes a first automated process control loop, the first automated process control loop comprising a first vessel, the first vessel comprising an inlet port and an outlet port, the inlet port coupled to a peristaltic pump to circulate a solution through a first sterilizing grade test filter, the first sterilizing grade test filter coupled to a first recovery filter, and the outlet port operable to receive filtered contents from the first recovery filter. The automated bacterial challenge test skid includes a second automated process control loop, the second automated process control loop comprising a second vessel, the second vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a second sterilizing grade test filter, the second sterilizing grade test filter coupled to a second recovery filter, and the outlet port operable to receive filtered contents from the second recovery filter. The automated bacterial challenge test skid includes a third automated process control loop, the third automated process control loop comprising a third vessel, the third vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a third sterilizing grade test filter, the third sterilizing grade test filter coupled to a third recovery filter, and the outlet port operable to receive filtered contents from the third recovery filter. The automated bacterial challenge test skid includes a fourth automated process control loop, the fourth automated process control loop comprising a fourth vessel, the fourth vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a fourth sterilizing grade test filter, the fourth sterilizing grade test filter coupled to a fourth recovery filter, and the outlet port operable to receive filtered contents from the fourth recovery filter, wherein the first automated process control loop in conjunction with the second automated process control loop, the third automated process control loop, and the fourth automated process control loop prevents test invalidation owing to failure of a single filter.

[7] An automated bacterial challenge test skid for open source validation and operable with a plurality of filter-make includes a first automated process control loop, the first automated process control loop comprising a first vessel, the first vessel comprising an inlet port and an outlet port, the inlet port coupled to a peristaltic pump to circulate a solution through a first sterilizing grade test filter, the first sterilizing grade test filter coupled to a first recovery filter, and the outlet port operable to receive filtered contents from the first recovery filter. The automated bacterial recovery test skid includes a second automated process control loop, the second automated process control loop comprising a second vessel, the second vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a second sterilizing grade test filter, the second sterilizing grade test filter coupled to a second recovery filter, and the outlet port operable to receive filtered contents from the second recovery filter. The automated bacterial challenge test skid includes a third automated process control loop, the third automated process control loop includes a third vessel, the third vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a third sterilizing grade test filter, the third sterilizing grade test filter coupled to a third recovery filter, and the outlet port operable to receive filtered contents from the third recovery filter. The automated bacterial challenge test skid includes a fourth automated process control loop, the fourth automated process control loop comprising a fourth vessel, the fourth vessel comprising an inlet port and an outlet port, the inlet port coupled to the peristaltic pump to circulate a solution through a fourth sterilizing grade test filter, the fourth sterilizing grade test filter coupled to a fourth recovery filter, and the outlet port operable to receive filtered contents from the fourth recovery filter, wherein the first automated process control loop in conjunction with the second automated process control loop, the third automated process control loop, and the fourth automated process control loop allows validation of one or more sterilizing filter-make provided by a customer for bacterial retention capability.

[8] The features and advantages described in this summary and in the following detailed description are not all-inclusive, and particularly, many additional features and advantages will be apparent to one of ordinary skill in the relevant art in view of the drawings, specification, and claims hereof. Moreover, it should be noted that the language used in the specification has been principally selected for readability and instructional purposes, and may not have been selected to delineate or circumscribe the inventive subject matter, resort to the claims being necessary to determine such inventive subject matter.

BRIEF DESCRIPTION OF FIGURES

[9] In the following drawings like reference numbers are used to refer to like elements. Although the following figures depict various examples of the invention, the invention is not limited to the examples depicted in the figures.

[10] FIG. 1A is a process flow diagram in accordance with one embodiment of the present invention;

[11] FIG. 1B is a process flow diagram in accordance with another embodiment of the present invention; and

[12] FIG. 2 is a system for preventing test invalidation owing to failure of a single filter, in accordance with one embodiment.

DETAILED DESCRIPTION

[13] In the present disclosure, relational terms such as first and second, and the like, may be used to distinguish one entity from the other, without necessarily implying any actual relationship or order between such entities.

[14] The following description outlines various embodiments of an automated bacterial challenge test rig that can be used for filter validation by any third party lab. The third party lab is equipped to supply process utilities like clean steam, air etc. The bacterial challenge test skid is on a mobile skid and can be taken from one place to another. In some instances, the bacterial challenge test skid can be installed at a customer’s premise also for demonstration purpose. The bacteria along with the drug product suspension are transferred to more than one vessel in the bacterial challenge test skid. Typically, the drug product is spiked with Brevundimonas diminuta (ATCC 19146) at a concentration =1x107 CFU/cm2 of effective filtration area after being proven that the viability of the bacteria is not compromised when in product or use a surrogate fluid for the challenge part of the test. A set of filters is used in the bacterial challenge test. The set of filters include a test filter, i.e. the filter to be tested for bacterial retention, and a recovery filter, i.e. the filter to recover any bacteria if the test filter fails to retain the bacteria. Typically, the recovery filter will be 0.45 micron rated filter membrane and the test filter will be a sterilizing grade test filter of 0.2 or 0.22 micron rating.

[15] FIG. 1A and FIG. 1B are process flow diagrams depicting various stages carried out in the automated bacterial retention skid. Before carrying out a Bacterial Retention Test, the first step is to carry out a viability test of the bacteria Brevundimonas diminuta (ATCC 19146) under process conditions (Temperature and contact time) with the product. If there is less than 1-Log reduction after the viability test, then the bacteria is considered to survive in the product under process conditions. FIG. 1A depicts the process flow diagram carried out if there is less than 1-Log reduction after the viability test. FIG. 1B depicts the process flow diagram carried out if there is more than 1-Log reduction after the viability test.

[16] Referring to FIG. 1A now:

[17] At step 105, an automated Clean-in-Place (CIP) is performed. CIP cleans the interior surfaces of pipes, vessels, process equipment, and associated fittings, without disassembly. CIP is usually carried out with one of purified water, acid, alkali, water-for-injection, or a combination thereof.

[18] At step 110, a Sterilization-in-place (SIP) is performed. SIP sterilizes the interior surfaces of pipes, vessels, process equipment, and associated fittings, without disassembly. SIP is usually carried out by passing clean steam. The test filters shall be bubble point tested prior to sterilization for integrity.

[19] At step 115, at the end of the viability test, the bacteria Brevundimonas diminuta (ATCC 19146) is directly inoculated into the product and is recirculated.

[20] At step 120, CIP is repeated once again.

[21] Referring to FIG. 1B now:

[22] The process steps depicted in FIG. 1B are carried out when the Log Reduction is more than 1 in the presence of the product under process conditions.

[23] Step 125 is similar to step 105. Likewise step 130 is similar to step 110.

[24] At step 135, the test filters are pre-conditioned with the drug product by recirculating as per the scaled-down simulated process conditions.

[25] At step 140, the test filters are flushed with the flushing fluids as per the validated flushing procedure to remove the residual product having antimicrobial effect. The flushing agents are recirculated and drained.

[26] At step 145, the automated bacterial retention skid is prepared for the bacterial challenge test after filter pre-conditioning and adequate rinsing. A surrogate fluid, for example, Saline Lactose Broth is inoculated with bacteria and further filtration is performed as per the scaled-down simulated process conditions, under bacterial challenge test.

[27] At step 150, the surrogate fluid containing the bacteria is recirculated.

[28] At step 155, CIP is once again carried out in the automated bacterial retention skid.

[29] The various steps along with functionality of various loops and associated process equipment, filters, and associated fittings, are explained in detail in conjunction with FIG. 2. Further, it is to be noted that the above mentioned steps are automated and controlled through a human machine interface or a SCADA.

[30] Referring to FIG. 2 now, an automated bacterial challenge test skid 200 for preventing test invalidation owing to failure of a single filter includes a first automated process control loop 202a. The first automated process control loop 202a includes a first vessel 210a. The first vessel 210a includes an inlet port 212a and an outlet port 214a. The inlet port 212a is coupled to a peristaltic pump 216 to circulate a solution through a first sterilizing grade test filter 226a. The first sterilizing grade test filter 226a is coupled to a first recovery filter 228a. The outlet port 214a is operable to receive filtered contents from the first recovery filter 228a.

[31] The automated bacterial recovery test skid 200 includes a second automated process control loop 202b. The second automated process control loop 202b includes a second vessel 210b. The second vessel 210b includes an inlet port 212b and an outlet port 214b. The inlet port 212b is coupled to the peristaltic pump 216 to circulate a solution through a second sterilizing grade test filter 226b. The second sterilizing grade test filter 226b is coupled to a second recovery filter 228b. The outlet port 214b is operable to receive filtered contents from the second recovery filter 228b.

[32] The automated bacterial challenge test skid 200 includes a third automated process control loop 202c. The third automated process control loop 202c includes a third vessel 210c. The third vessel 210c includes an inlet port 212c and an outlet port 214c. The inlet port 212c is coupled to the peristaltic pump 216 to circulate a solution through a third sterilizing grade test filter 226c. The third sterilizing grade test filter 226c is coupled to a third recovery filter 228c. The outlet port 214c is operable to receive filtered contents from the third recovery filter 228c.

[33] The automated bacterial challenge test skid 200 includes a fourth automated process control loop 202d. The fourth automated process control loop 202d includes a fourth vessel 210d. The fourth vessel 210d includes an inlet port 212d and an outlet port 214d. The inlet port 212d is coupled to the peristaltic pump 216 to circulate a solution through a fourth sterilizing grade test filter 226d. The fourth sterilizing grade test filter 226d is coupled to a fourth recovery filter 228d. The outlet port 214d is operable to receive filtered contents from the fourth recovery filter 228d, wherein the first automated process control loop 202a in conjunction with the second automated process control loop 202b, the third automated process control loop 202c, and the fourth automated process control loop 202d prevents test invalidation owing to failure of a single filter.

[34] Further the first automated process control loop 202a in conjunction with the second automated process control loop 202b, the third automated process control loop 202c, and the fourth automated process control loop 202d prevents test invalidation owing to contamination of a single vessel. The automated bacterial challenge test skid 200 includes a cryo-bath 236 for maintaining an even distribution of temperature across one or more vessels. The cryo-bath 236 includes a temperature transmitter 234. The first automated process control loop 202a, the second automated process control loop 202b, the third automated process control loop 202c, and the fourth automated process control loop 202d enable automatic control of one or more process parameters. The one or more process parameters include at least one of temperature, pressure, differential pressure, contact time, contact time, flow rate, and total volume filtered. A change in the one or more process parameters are captured to provide a 21 CFR part 11 compliant audit trail. The automated bacterial challenge test skid 200 is operable with a plurality of filter-make. The automated bacterial challenge test skid 200 is configured such a way that it is suitable for open source validation, where any sterilizing filter of a customer’s choice can be validated for its bacterial retention capability. This gives the customers a truly selected and optimized filtration train.

Clean-in-place (CIP)
[35] The sterilizing grade test filters and the retention filters are removed during CIP and instead a spool piece (not shown in figure) is connected. A flushing liquid is added into the vessels 210a, 210b, 210c, and 210d through the respective pump head of the pump 216. Further, the respective outlet tubes of outlet ports 214a, 214b, 214c, and 214d are coupled to drain 274 via manual valves 204a, 204b, 204c, and 204d. The drain valves 206a, 206b, 206c, and 206d are open. Then flushing is started. After flushing, the outlet tubes are connected back to respective vessels and the drain valves 206a, 206b, 206c, and 206d are closed. The alkali is added into the vessel through the respective pump head of the pump 216 and recirculated for a set time. Further, it is drained out by opening the valves 208a, 208b, 208c, and 208d as well as the drain valves 206a, 206b, 206c, and 206d. Further purified water is added into the vessel through the peristaltic pump 216and recirculated for a set time. Further, it is drained out by opening the valves 208a, 208b, 208c, and 208d as well as 206a, 206b, 206c, and 206d. Further acid is added into the vessel through the peristaltic pump 216 and recirculated for a set time. Further, it is drained out by opening the valves 208a, 208b, 208c, and 208d as well as 206a, 206b, 206c, and 206d. Further purified water is added into the vessel through the peristaltic pump 216 and recirculated for a set time. Further, it is drained out by opening the valves 208a, 208b, 208c, and 208d as well as 206a, 206b, 206c, and 206d. Further purified water or water for injection is added into the vessel through the peristaltic pump216 and recirculated for a set time. Further, it is drained out by opening the valves 208a, 208b, 208c, and 208d as well as 206a, 206b, 206c, and 206d. The cycle is continued till conductivity reaches set value. Once conductivity levels are reached air is flushed by opening valves 208a, 208b, 208c, and 208d as well as 220a, 220b, 220c, and 220d. Further, manual valves 204a, 204b, 204c, and 204d are closed.

Sterilization-in-place (SIP)
[36] The sterilizing grade test filters are removed during SIP as well as the flexible hoses are removed. Dummies are put in inlet hose. The spool piece is connected and the vent filters are clamped. Clean steam is brought into the skid 200 by opening manual utility valve 240. The pressure of the clean steam is maintained at the set point by using the pressure regulating valve 242. The pressure of steam is indicated in 244. The steam is controlled by valve 246 to maintain the temperature at set point. The temperature transmitter 258, 260, 262, and 264 provides a feedback for controlling the temperature of the steam at the set point. The non-return valves 266, 268, 270, and 272 prevent back-flow of any liquid into the skid 200.

[37] The steam can be passed into the skid 200 by opening valves 218a, 218b, 218c, and 218d as well as valves 208a, 208b, 208c, and 208d. Likewise, process air utility is brought into the skid by opening manual utility valve 250. The pressure of air is maintained at the set point by using the pressure regulating valve 252. The air is filtered through a hydrophobic air filter 254. Air inlet is controlled by opening or closing the actuated valve 256. The manual valves 204a, 204b, 204c, and 204d are crack open till a set point temperature is reached. Sterilization is carried out for the determined period. Then 204a, 204b, 204c, and 204d are closed followed by closing of drain valves 206a, 206b, 206c, and 206d. This is followed by cooling of the vessel by opening the valves 220a, 220b, 220c, and 220d as well as 256.

Recirculation
[38] After CIP and SIP are completed sterilizing grade test filters and the retention filters as well as the inlet hoses are connected and the vent filters are de-clamped. The media is recirculated at a set flow rate indicated by the flow indicator-controller-recorder 222a, 222b, 222c, and 222d and controlled by the speed indicator-controller-recorder 232 of peristaltic pump 216. Manual valves 204a, 204b, 204c, and 204d are opened and a collection beaker is placed. Collection fluid or the filtrate is collected by opening actuated valves 208a, 208b, 208c, and 208d. Manual valves 204a, 204b, 204c, and 204d are closed. Recirculation can be of two types. Continuous recirculation is where the step is completed after recirculation timer is elapsed. Second type of recirculation is intermittent recirculation where the recirculation is stopped and started at regular intervals as indicated in the Process Questionnaire. After the sequence, total volume filtered, cycle time, total contact time, temperature, pressure, differential pressure, and the flow rate are displayed on a HMI/SCADA screen.

Flushing
[39] A flushing or cleaning agent is added to the vessels. The manual valve 204a, 204b, 204c, and 204d are opened. The flushing agent is circulated through peristaltic pump216and controlled by speed indicator-controller-recorder 232. The pressure is controlled by a pressure indicator-controller-recorder 224a, 224b, 224c, 224d, 230a, 230b, 230c, and 230d. After recirculation time is completed the flushing agent is drained by opening actuated valves 208a, 208b, 208c, and 208d. When flow level shown is low, the manual valves 204a, 204b, 204c, and 204d are closed.

[40] There are many inherent advantages of the proposed skid. Firstly, use of the cryo-bath (temperature controller cabinet) provides even distribution of temperature across the vessels. Secondly, the minimum volume that can be used effectively is as low as 250 mL. The proposed skid provides efficient cleaning considering the small size. The cleaning time and solution required will also be very less. Since the skid includes individual control loops the flow distribution will be simple and efficient across the skid. Further, having 4 different vessels avoids cross-contamination.

[41] The following detailed description is intended to provide example implementations to one of ordinary skill in the art, and is not intended to limit the invention to the explicit disclosure, as one or ordinary skill in the art will understand that variations can be substituted that are within the scope of the invention as described. For example, the vessels which are of stainless steel make can be replaced with Cartridge or Capsule or disposable bags depending on the requirement of the customer. Also multiple lines or loops can be added to run two or three tests in parallel, depending on the test conditions, to enhance the productivity. Likewise, the skid can be used for performing mycoplasma retention studies employing suitable procedures. The mycoplasma retention test is carried out using the challenge organism “Acholeplasma spp”.

[42] The skid can be used for sterilizing grade as well as non-sterilizing grade filters for its bacterial retention capability. The applications of the skid include but are not limited to biotech industry, water purification, pharmaceutical, diagnostic kits manufacturing, and breweries. The skid can be used for validation of sterile filtration of fluids, air, and process gases. The skid can be used by testing labs, filter manufacturers, third party testing labs, contract research organizations and academic institutions. The skid can be used for the validation of different dosage forms including injectable solutions, intravenous injections, nasal sprays and all small and large volume parenteral, novel drug delivery systems including liposomes, microspheres, and delayed and sustained release forms.

[43] In the preceding specification, the present disclosure and its advantages have been described with reference to specific embodiments. However, it will be apparent to a person of ordinary skill in the art that various modifications and changes can be made, without departing from the scope of the present disclosure, as set forth in the claims below. Accordingly, the specification and figures are to be regarded as illustrative examples of the present disclosure, rather than in restrictive sense. All such possible modifications are intended to be included within the scope of present disclosure.