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Technical Summary
Polyclonal antibody production is performed using pathogen-free, barrier-reared animals. Barrier raised animals provide greater reproducibility and often produce higher titres with lower background.
Antigens
Antigen can be supplied as either in solution, lyophilized or conjugation to a carrier protein. We also offer conjugation of peptide to a carrier service.
Typical antigen concentrations required for each immunization each time are listed blow.
| Rabbit |
Goat |
Rat |
Mouse |
| 50-200µg |
>200µg |
10-25µg |
5-20µg |
Facility and Process
Our facilities are regulated and controlled under strict Home Office Guidelines ensuring a high standard of animal care and documentation by dedicated staff. Standard operating procedures ( SOPs ) are in place when performing each process, i.e. Immunization SOP, Blood Sampling SOP and Serum Preparation SOP. All process records and data are fully documented allowing complete traceability.
Protocols
GeneMay will use established protocols for polyclonal antibody production in normal case. Alternatively, we can use customized protocols. Simply telling us the number of immunizations and bleeds you require, we will incorporate them into our standard protocol. We will then execute the procedure upon your approval. A protocol may be extended beyond it's original finish date. We will communicate with you to confirm by fax or e-mail in advance of the termination of animals. In order to meet our Home Office regulations, a brief justification of the project to be undertaken is required before the protocol can commence.
Animals
GeneMay can perform polyclonal antibody production in a variety of different species. Currently, Our service includs rabbit, Goat, Rat and Mouse.
Immunizations
Immunizations are performed according to the schedule by subcutaneous injection. Rodents can be immunized with up to 0.1ml at each of two sites giving a total volume per immunization of 0.2ml. Other animals may be immunized with up to 0.25ml at each of four sites giving a total volume of 1.0ml per immunization.
Adjuvants
There are a variety of adjuvants available for use in polyclonal antibody production.
Bleeds
Although no guarantee can be given regarding the final yield, typical volumes for antisera are listed below.
| |
Rabbit |
Goat |
Rat |
Mouse |
| Test Bleed |
2.5-5ml |
5-10ml |
0.5-1.0ml |
up to 0.2ml |
| Production Bleed |
up to 10ml |
up to 200ml |
N/A |
N/A |
| Terminal Bleed |
50-75ml |
500-1000ml |
5-10ml |
up to 2.5ml |
Test samples and production bleeds will be sent out within one week after colleciton unless further processing has been requested in advance.
Testing
GeneMay will perform ELISA against Ag tests for antibody titres for you. We also can perform a specific assay for your antigen if requested.
Purification
Antisera will be purified by either protein G purification or immunoaffinity resin using your ligand of choice.
Conjugation
GeneMay also offer conjugation service on your antibody. We currently offer the Ab conjugation service with FITC, PE, PE-Cy5, APC, Biotin, AKP and HRP.
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