Polyclonal Antibody Development
Polyclonal antibodies (pAbs), a complex mixture of many antibodies, are frequently produced by a variety of B-cell clones in the body. These antibodies can recognise and bind to several epitopes on a single antigen. Due to their multi-epitope binding properties, polyclonal antibodies are perfectly suited for many applications.
Fagus provides full-service packages for the manufacturing of polyclonal antibodies. We can a variety of service packages to accommodate your requirements. We have extensive experience in generating polyclonal antibodies.
Our standard package include:
- Conjugation of peptide or hapten to a carrier (If applicable), Immunisations, collection of pre-immune, test and terminal bleeds.
- Antibody titre or specificity testing.
- Serum purification – purification of the immunoglobulin can be undertaken by several methods
o Protein A/G purification (20-30 mls serum)
o Ion-exchange purification (20-30 mls serum)
o Peptide affinity purification (free N-terminus)
- Peptide affinity purification (Terminal cysteine)
Pricing dependent upon number of hosts and purification required – please enquire.
Timescale & Processes
Step 1 – Variable – Submission of justification for antibody generation for ethical review.
Step 2 – 2 weeks – Generation and/or coupling of peptide to BSA and KLH.
Step 3 – Immunisation protocol starts with pre-immune bleed.
Step 4 – Test bleed with ELISA.
Step 5 – 10 weeks from immunisation – Harvest and serum preparation with ELISA.
Step 6 – 1 week – Purification of e.g. 10 ml of serum (approximately 50 mg of total IgG) by Protein G
chromatography. Buffer exchange to required format, purity assessment by SDS PAGE and ELISA.
Step 7 – 2 weeks – Affinity chromatography from total IgG using coupled antigen. Buffer exchange to required
format, purity assessment by SDS PAGE and ELISA.