Veterinary Biologics Guideline 3.22E
Guideline for the Submission, Testing and Reporting of Veterinary Biological Samples

Table of Contents

• I. Introduction
• II. Legal authority
• III. Background information
• IV. Procedures for submitting master or working seeds and cell line samples
• V. Requirements for master seed bacteria
• VI. Requirements for master seed viruses
• VII. Requirements for master seed cell lines
• VIII. Requirements for pre and post-licensing serial testing
• IX. For further information

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I. Introduction

The purpose of this guideline is to notify manufacturers and importers of the requirements for the submission of veterinary biological samples to the Biologics Evaluation Laboratory (BEL), Canadian Food Inspection Agency (CFIA) or its designated laboratory facilities for quality assurance monitoring.

II. Legal authority

Health of Animals Regulations

Section 129. (1)
Every holder of a product licence shall
(a) keep and make available for inspection by a veterinary inspector records with respect to the preparation, manufacture, preservation, storage, testing, sale and distribution of every veterinary biologic manufactured under the product licence and any diluent to be used therewith.

Section 130.
No person shall sell a veterinary biologic or any diluent to be used therewith if the veterinary biologic or diluent has been prepared, manufactured, preserved, packed, labelled or tested otherwise than in the manner described in the product outline.

Section 131.
Where the Minister is satisfied from tests of a veterinary biologic, or otherwise, that a veterinary biologic is unsafe to use, is likely to cause communicable disease in animals or is contaminated or ineffective, he may, by order, prohibit the importation, manufacture, sale or distribution of the veterinary biologic.

III. Background information

A. The Canadian Centre for Veterinary Biologics (CCVB), Animal Health Division (AHD), CFIA may request that the following types of veterinary biologics samples be submitted to the BEL for quality assurance testing:

1. Master or working seed bacterial or viral stock
2. Master or working seed cell line
3. Pre-licensing serial
4. Post-licensing serial

B. The BEL will carry out master seed testing, pre-licensing serial testing and post-licensing serial release testing for veterinary biologics manufactured in authorized facilities in Canada or in foreign countries other than the United States (US).

C. The BEL does not usually test veterinary biologics manufactured in authorized facilities in the US, as these activities are done by the Center for Veterinary Biologics (CVB) of the United States Department of Agriculture (USDA).

1. Exception: A veterinary biologic manufactured in the US under the Food and Drugs Administration Export Reform and Enhancement Act of 1996 (FDA-EREA) will undergo master seed, pre-licensing serial and post-licensing serial testing as in IV.A.

D. The term "test", unless specifically stated otherwise, generally refers to tests for purity, potency, and/or safety.

E. Unless otherwise directed, all veterinary biologics samples are to be sent to the following address:

IV. Procedures for submitting master or working seeds and cell line samples

A. During the course of the review process, the CCVB reviewer responsible for the product submission will instruct the manufacturer to submit seed and/or cell line stock samples for laboratory evaluation and, at the same time, will advise the BEL of a pending submission.

B. Prior to submitting seed and cell line samples to the BEL, the manufacturer will contact the BEL to confirm shipping dates and sample and information requirements.

1. Samples for import into Canada will require a valid import permit issued by the CFIA-CCVB to accompany the shipment.

C. All shipments to the BEL must be accompanied by a cover letter clearly identifying the sample, the lot/serial number, the volume and number of doses, and the reconstitution directions.

1. The vial label information must match the information provided in the cover letter as well as that in the Outline of Production and other referenced documents.

D. The samples should be shipped in a manner that ensures maintenance of the recommended storage temperature in transit.

1. Note: Seed viral stocks and seed cell line stocks are to be shipped frozen on dry ice.

E. The BEL will notify the manufacturer and the CCVB of sample receipt. Individual samples will be inspected and deemed to be in either satisfactory or unsatisfactory condition upon arrival. Replacement samples will be requested for those samples arriving in an unsatisfactory condition.

F. All working seeds and cell lines, or master seeds and cell lines if they are used as working stock, shall be tested.

G. If, at any time, a manufacturer intends to put into use a new master or working seed or cell line, the manufacturer is required to inform CCVB and the new stock(s) should be resubmitted to the BEL (as above) for testing.

H. The following examples of problems encountered with sample submission to the BEL will result in delays to sample testing:

1. Label information not matching the cover letter information
2. Illegible or missing vial labels
3. Lack of a cover letter
4. Volume and/or diluent and/or reconstitution directions not provided
5. No or incomplete supporting information as specified in sections V.B., VI.C., and VII.C below
6. Insufficient sample volume and/or vial numbers
7. Neutralizing antiserum and/or other assay specific reagents (i.e. reference standards) as specified in the Outline of Production not provided
8. Shipping temperature out of range

V. Requirements for master seed bacteria

A. Sample requirements:

1. For frozen cultures a minimum of 2 vials containing at least 1 mL per vial is required.
2. For lyophilized cultures at least 2 vials are required
3. For bacterial seeds derived from biotechnology an additional vial is required.

B. Prior to the shipment of master seed bacteria and to facilitate laboratory testing the manufacturer is required to provide the following information to the BEL in advance of sample submission:

1. Genus, species and additional taxonomic data (e.g. serotype if available, description of genetic modification if applicable).
2. Origin and history
3. Conditions for storage (e.g. media, temperature, lyophilized, frozen etc.)
4. Conditions for propagation (e.g. media, temperature, CO₂, aeration etc.)
5. Results of any characterization and identity testing (e.g. morphological characteristics, temperature range of growth, speciation by biochemical testing, serotyping etc.)

VI. Requirements for master seed viruses

A. Samples of each virus from the maximum allowable passage level for vaccine production (as defined in the Outline of Production) shall be tested. Passaging of virus is to be carried out by the manufacturer. The virus may be additionally passaged by the BEL.

B. Sample requirements

1. If the manufacturer has separate master and working virus seeds, a minimum of 2 vials with a minimum total volume of 2 mL of the former and a minimum of 4 vials with a minimum total volume of 4 mL of the latter are required.
2. If master seed and working virus seeds are one and the same, a minimum of 6 vials with a minimum total volume of 6 mL are required.
3. If available, a minimum of 2 vials with a minimum total volume of 2 mL of neutralizing antiserum to master seed virus (if virus was totally neutralized in a maximum 1:1 ratio with antiserum) would also be required.
4. Note: For genetically modified viruses, an additional one mL of each sample type (master and/or working seed, antiserum) is required.

C. Prior to the shipment of master seed viruses and to facilitate laboratory testing the manufacturer should provide the following information to the BEL:

1. History
   a. name (family, genus) of virus
   b. origin of virus
   c. source of virus
   d. passage history
   e. maximum virus passage allowable for vaccine production lot
   f. titre of master seed
2. Growth Conditions
   a. cell line(s) generally used
   b. medium, incubation temperature
   c. multiplicity of infection, adsorption time, usual time of harvest
   d. expected titre range
3. Storage Conditions
   a. temperature
   b. condition (liquid or lyophilized)
   c. stability
4. Results of any quality control testing done

VII. Requirements for master seed cell lines

A. Samples of each production lot derived from master or working seed banks as described above shall be tested at ≥ 5 passages beyond the maximum allowable passage as defined in the Outline of Production.

1. Passaging of cells is to be carried out by the manufacturer. Cells may also be passaged additionally by the BEL.

B. Sample requirements

1. If the manufacturer has separate master and working cell stocks, a minimum of 2 vials with a minimum total volume of 2 mL of master stock and a minimum of 4 vials with a minimum total volume of 4 mL of working stock are required.
2. If master and working cell stocks are one and the same, a minimum of 6 vials with a minimum total volume of 6 mL are required.

C. Prior to the shipment of master seed cell lines and to facilitate laboratory testing the manufacturer should provide the following information to the BEL:

1. History
   a. identification (name and species of cell line)
   b. origin of cell line
   c. source of cell line
   d. passage history of cells
   e. maximum cell passage allowable for vaccine production lot
2. Growth conditions
   a. normal split ratio
   b. growth medium
   c. temperature, CO₂, humidity requirements
   d. maintenance medium
3. Freezing conditions
   a. freeze medium
   b. cell density recommended
   c. conditions (liquid or vapour phase)
4. Results of any quality control testing done

VIII. Requirements for pre and post licensing serial testing

A. Testing of a serial or lot can be one or any combination of tests on that product or serial.

B. The first three pre-licensing serials of any product shall be tested.

1. Rates of testing of subsequent pre-licensing serials, if required, will be determined by the CCVB reviewer in consultation with the BEL.

C. The first three post-licensing serials of any product shall be tested.

1. Subsequent post-licensing serials shall be tested at the rate of approximately twenty percent of the total number of serials of any given product received (based on previous numbers received). A product will be tested at least once in a 12 month period when there are fewer than five submissions during the same period.
2. Products that have a history of problems (e.g. contamination, adverse reactions), or have been produced according to modifications or changes in the Outline of Production, or for other appropriate reasons, may be tested more frequently.

D. Sub-serials will be considered as separate serials for testing purposes, for all categories of testing, i.e. purity, potency, and safety.

1. All sub-serials of any given product, when received concurrently, will not be released until the results of any tested sub-serials are known.
2. If failure occurs in one or more sub-serials of a product, the release of all sub-serials may be re-assessed.

E. To avoid unnecessary delay in releasing the test results, samples of each product should be submitted to the BEL when final container filling is completed.

1. The manufacturer should wait for instruction from the CCVB reviewer before submission of pre-licensing serials to the BEL.

F. The requirements for the number of vials or containers to be submitted for each serial are listed below:

BEL Sample Requirement
The following number of vials per serial must be submitted for routine testing:

1. All products except those listed below.

• 10 vials of Size < 10 mL
• 8 vials of Size 10-20
• 3 vials of Size 21-100
• 1 vial of Size >100 mL

2. Diagnostic kits

• minimum 2 kits

3. Colostral whey products

• minimum 2 doses

4. Antigens

• minimum 3 mL

5. Mannheimia haemolytica and Haemophilus somnus containing bacterins or extracts (bovine only)

• in addition to the vials required in 1. above, an equivalent amount of pre-adjuvanted antigen is to be submitted

Note: An equivalent number of sample units are to be retained by the manufacturer or importer for a minimum of 6 months past the expiration date, should a retest by the BEL be required.

G. All shipments to the BEL must be accompanied by a cover letter clearly identifying the sample, the lot/serial number, the volume and number of doses, and the expiration date.

1. Each container or vial will have an approved label containing, as a minimum, the information outlined in CCVB Guideline 3.3 - Guideline for labelling of veterinary biologics.
2. Each submission will be sent in the approved product carton, accompanied by the insert and the recommended diluent, where applicable
3. The vial label information must match the information provided in the cover letter as well as that in the approved Outline of Production on file at CCVB.

H. The samples should be shipped in a manner that ensures maintenance of the recommended storage temperature in transit.

I. The BEL will notify the manufacturer and the CCVB of sample receipt. Individual samples will be inspected and deemed to be in either satisfactory or unsatisfactory condition upon arrival. Replacement samples will be requested for those samples arriving in an unsatisfactory condition.

J. All manufacturers' test results for each serial are to be forwarded to the CCVB for approval and release of the serial (using the form Manufacturer's Serial Release Test Report).

1. The test references on the serial release test result forms must cite the current Outline of Production and referenced Special Outlines as filed at CCVB.
2. The Manufacturer's Serial Release Test Report must be accompanied by an Application for Services (Form CFIA/ACIA 4720)
3. The manufacturers are requested not to send test reports or fees to the BEL.

K. Upon receipt of both the manufacturer's serial test results and the BEL serial test report by CCVB, the authority to release the serial will be sent by CCVB directly to the manufacturer by the reviewer in charge of serial release.

1. Should the tests conducted yield suspicious, equivocal or results interpreted as test failure, the manufacturer will be notified and the tests may be repeated.
2. If the serial is not selected for testing by the BEL, the authority to release the serial will be sent to the manufacturer upon receipt of the manufacturer's serial release test results by CCVB.

L. Note: In special cases (i.e. diagnostic kit testing in support of CFIA programs), serial testing will be carried out by another CFIA testing facility. In these instances, the CCVB reviewer will provide additional guidance to the manufacturer with regards to the shipping of samples. Similar requirements to those described above will apply.

IX. For further information

Any further questions pertaining to veterinary biological samples may be directed to Canadian Centre for Veterinary Biologics.