Q and A Neutralization Assay A002 final.pdf

Question Government Response

As it relates to the above subject solicitation, can the government confirm that it already has a qualified SARS-CoV-2 virus specific neutralization assay?

The laboratory work has been completed and the statistical analysis is ongoing. A qualification report is expected by 8/7/2020.

Will BARDA provide the SARS-CoV-2 assay and validation protocols or does BARDA expect the Contractor to propose an assay and validation protocols?

BARDA will provide a technology transfer protocol which will include the following: i. An assay specific Standard Operating Procedures (SOP)

ii. A qualification protocol.

iii. A plan for stability testing of reagents

iv. Comparability panel

v. A qualification report The Offeror shall provide:

i. A validation protocol that evaluates precision, dilutional linearity, LOD, LOQ, selectivity (matrix effects), and specificity as appropriate. The validation protocol should include a statistical analysis plan.

ii. A plan for stability testing of reagents

iii. A validation report

Are there a number of assays and time period? That would be helpful in providing a ROM price?

Offerors proposing less than 1000 samples per week (40 hours/week) will not be considered responsive. The proposed immunoassay capacity must be available for use in full no later than October 30, 2020. The total proposed duration of this work is 12 months from award.

Is compliance with GLP acceptable in lieu of GCLP? Yes For the ROM, do you have a rough sample size in mind for the LOE?

Asking because the range could be vast when looking at say the current sample group of 30,000 receiving vaccine trials.

Offerors proposing less than 1000 samples per week (40 hours/week) will not be considered responsive. The proposed immunoassay capacity must be available for use in full no later than October 30, 2020. The total proposed duration of this work is 12 months from award.

Can you share the SOP and how detailed is it? SOP cannot be shared at present.

If the SOP cannot be shared at present, we have the following questions:

1. What is the method of detection of viral replication? - CPE, qPCR for viral RNA, qPCR for mRNA, immunostaining, others? This would affect the facilities/instruments needed, the duration of the assay and the number of personnel needed

The read outs will be CPE and ELISA .

What is the preferred range and number of dilutions - for a range of 1:10 to 1:1000, the number of dilutions can be 3 for log dilutions, 4-5 for 5-fold dilutions and 8 for 2-fold dilutions

Range is 1:20 – 1: 2560. Initial 1:20 followed by 7 2-fold dilutions. For higher titer samples starting dilution can be increased.

What is the expected number of replicates per dilution - minimum would be 4 but higher the number of replicates, more accurate the assay

There are two replicates per dilution

What is the preferred format of the assay - 24 well, 96 well or 384 or 1,536 wells? That will determine the number of samples per plate. For 8 dilutions at 4 wells/dilution (i.e. 32 wells/sample) the number of plates for 1000 samples will be: 96 wells - 250; 384 wells - 84; 1,536 wells - 28

A 96-well plate is used.

5. Will there be any gold standard samples provided with known concentrations of neutralizing antibodies that can allow us to quantify the mass concentrations of the neutralizing antibodies?

Not yet, preparation in progress.

Is the ultimate intent of the assay an exact quantification of the antibody response or an yes/no answer. The yes/no level would depend on the established MDLs (Medical decision levels)

Purpose is quantification of antibody titer

Can you clarify what assays will be tech transferred and validated? The announcement is for live virus neutralization (only, no other assays)tech transfer, validation and testing of Ph3 COVID-19 vaccine trial samples.

Under task 5 it references cGMP regulations but I don’t think these regulations apply as nothing is being manufactured (just testing samples)

N/A

Would it be acceptable to respond that samples would be tested under

GLP?

Yes, GLP is acceptable.

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