APMA 2008CBER Reference/Sera

Candidacy Testing

September 2008

James King

9/04/03

Agenda

Review of last years commitment to CBER Proposal for standardizing the optimization

process for reference or sera replacement Proposal for summarizing reference

replacement and unknown ERP data Discuss minimum aliquots required for

proposed testing of reference or sera replacements

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APMA meeting October 2007

Industry accepted the need to partner with CBER to provide quick and useful data for reference and sera replacement

Industry committed to have reference review to CBER in 30 days

Per the meeting minutes, CBER was going to supply industry with a standard method to supply results

CBER or Industry to standardize method for communicating reference/sera review

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To facilitate a unified collaboration between industry and CBER, HollisterStier Laboratories would like to propose:

A systematic approach to review new reference or sera

A means to report the data in a standardized format

Sharing data between industry and CBER

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Approach for replacement review

Optimization of assay parameters

Side by side testing of approved and proposed system

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Optimization

Goals:– Perform checkerboard optimization to achieve

desired signal to noise.– Minimize consumption of CBER reference

and sera.– Formalize a standard practice to communicate

results to CBER.

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Optimization Proposal

Product Name = Mite D. Farinae Coating Dilutions

1:750 1:100 1:250 1:500 1:750 1:1000 1:750 1:100 1:250 1:500 1:750 1:10001 (Blank) 2 3 4 5 6 7 (Blank) 8 9 10 11 12

Sera 1:5 ADilution 1:10 B High Control High Control

1:20 C Anti-IgE 1:1000 Anti-IgE 1:20001:25 D

HSA 1:5 EDilution 1:10 F Low Control Low Control

1:25 G Anti-IgE 1:1000 Anti-IgE 1:20001:50 H

Exsisting coating dilution no sera added Exsisting coating dilution no sera added

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Example Checkerboard

Product Name = Mite D. pteronyssinus Sera Optimization

BLANK 1:100 1:250 1:500 1:750 1:1000 BLANK 1:100 1:250 1:500 1:750 1:1000A 0.221 1.389 1.220 1.051 0.882 0.713 0.198 1.220 1.051 0.882 0.713 0.544B 0.200 0.987 0.890 0.793 0.696 0.599 0.190 0.890 0.793 0.696 0.599 0.502C 0.198 0.560 0.401 0.242 0.242 0.231 0.179 0.560 0.535 0.510 0.485 0.460D 0.210 0.301 0.270 0.239 0.208 0.177 0.182 0.230 0.277 0.324 0.371 0.418E 0.201 0.289 0.287 0.285 0.283 0.281 0.176 0.289 0.287 0.285 0.283 0.281F 0.189 0.270 0.267 0.264 0.261 0.258 0.198 0.270 0.267 0.264 0.261 0.258G 0.220 0.200 0.225 0.250 0.275 0.300 0.189 0.200 0.225 0.250 0.275 0.300H 0.225 0.198 0.210 0.222 0.234 0.246 0.190 0.198 0.210 0.222 0.234 0.246

Selected Conditions:Sera Dilution 1:20Coating Dilution 1:1000Anti IgE dilutuion 1:1000

Low Control (Anti IgE 1:2000)High control (Anti IgE 1:1000) Low Control (Anti IgE 1:1000)

High control (Anti IgE 1:2000)

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Performing CBER side by side testing

CBER distributes new reference – Two unknown samples are included– Third sample reserved for reference

comparison– Samples tested on current system – Samples are tested on new optimized

system

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Example of Results for submission to CBER and Industry

Test 1logRP RP logRP RP logRP RP logRP RP

Plate 1 0.0866 1.2207 -0.0321 0.9288 -0.0380 0.9163 0.1302 1.3494Plate 2 -0.0048 0.9861 -0.0707 0.8498 -0.0748 0.8418 -0.0629 0.8652Plate 3 0.1112 1.2919 -0.0410 0.9099 -0.0495 0.8922 -0.0399 0.9122Mean 0.0643 1.1597 -0.0479 0.8955 -0.0541 0.8829 0.0091 1.0213

Test 2logRP RP logRP RP logRP RP logRP RP

Plate 1 0.0289 1.0689 0.0079 1.0183 0.0172 1.0404 -0.0367 0.9189Plate 2 0.0858 1.2184 0.0321 1.0767 0.0494 1.1206 -0.0090 0.9795Plate 3 0.0938 1.2412 0.0171 1.0402 0.0429 1.1039 -0.0466 0.8982Mean 0.0695 1.1735 0.0190 1.0448 0.0365 1.0877 -0.0308 0.9316

Percent Difference 1% 15% 20% 9%

E9-Df as Ref Std and Coating Extract (1:500)CBER Sample #1 CBER Sample #2 E9-Df QC RefE8-Df

E8-Df QC RefE9-DfE8-Df as Ref Std and Coating Extract (1:500)

CBER Sample #1 CBER Sample #2

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HollisterStier will provide to those interested:

Spreadsheet for checkerboard optimization– Including fields for identifying selected

conditions Spreadsheet for side by side comparisons

– Including automated calculation for percent difference or other statistical method as discussed by CBER and industry

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Needs from CBER

Provide enough of the proposed reference or sera to perform the testing as outlined in this slide show.

– Approximately 500µL reference needed– Approximately 2 or 3 vials of sera

Provide unknown test samples for comparative testing

Communicate with industry when references are low so we can better adjust production

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In summary

This process is intended to:– expedite testing of proposed CBER

reference or sera replacements– minimize impact to industry– summarize and share test results with

industry in a standardized format– provide justification for accepting or

rejecting reference or serum replacement– most importantly, allow us to work

together

9/04/03

Thank you for your time

Remember to submit your yearly forecast of CBER reference and sera consumption to

CBER by January 2009