Custom Polyclonal Antibodies

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Proteintech offers a robust immunization and serum collection protocol for custom rabbit polyclonal antibodies made from synthesized peptides or recombinant proteins. Proteintech’s insightful knowledge of antigen choice (specificity, affinity, protocol optimization) means we produce a high standard custom antibody tailored for your needs.

 

Peptide made polyclonal antibodies

 

Peptide antibodies can be designed to perform specifically for your intended application, or to maximize cross-reactivity in orthologous sequences. Our service includes; peptide design, synthesis, conjugation to KLH, immunization, and affinity purification.

 

For the peptide design, our antibody experts review the protein sequences provided by you and rank the most antigenic domain. We then submit a report to you to make the final selection. We aim to have this stage complete within 24 hours of receiving your inquiry.

 

The yield of the peptide synthesis is approximately 40 mg, with around 90% purity for 20 mer-peptides with Mass Spectrum data. Due to the high purity, there is no need for an extra affinity purification stage. If you require additional peptides to be synthesized, there will be an additional charge. This stage usually takes one week.

 

Additional peptide service available: Phospho-specific or acetylation specific peptide design.

 

Peptide Conjugation info: 10 mg of peptide is conjugated with 5 mg of KLH protein. We can also offer BSA at customer’s request.

 

Whole protein made polyclonal antibodies

Whole protein immunogens often make higher quality antibodies as they recognize a wider range of epitopes and have higher binding affinity. The protein antibody production service includes:

  • Cloning
  • Protein expression
  • Immunization
  • Affinity purification

Cloning and protein expression process:

  • Proteintech will design the most antigenic and divergent coding region of 300 amino acids or less to be the antigen.
  • PCR amplification of the desired coding region.
  • Subcloning the cDNA into any expression plasmid that the customer provided.
  • Large Scale culture, induction, and purification of 1 mg of the fusion protein.                                                    
  • This service is available without cloning if you provide the expression vector.

Affinity purification: The standard service includes affinity purification column preparation and 30ml of antiserum purification with the column. The customer will receive the column, purified antibody, flow through, and the remaining serum.We determine which rabbits’ antisera to purify by the ELISA result from the test bleed. If both rabbits give good results, we will purify 15ml of serum from each rabbit. If one rabbit gives better results than the other, we will purify 30ml of serum from this rabbit.

Depending on the concentration of the specific antibody in the serum provided we can usually get between 2-10ml from 15ml antiserum. For an additional charge, you can request more antisera to be affinity purified. 

Proteintech's 102-day immunization protocol

Day 0 Pre-immune bleed
Day 1 Primary injection
Day 28 Boost 1
Day 42 Boost 2
Day 56 Test bleed and ELISA
Day 60 Boost 3
Day 74 Production bleed
Day 78 Boost 4
Day 88 Production bleed
Day 102 Final bleed

This protocol is optimized to increase the titer and volume of the obtained antibody. The 102-day immunization ensures enough time for the antibody to mature. Each rabbit will give ~12 ml of serum in each production bleed and 45-50 ml of serum (from 90 - 100 ml blood) in the final bleed, a total of about 150 ml of antiserum from two rabbits.

Timescales

As custom antibody production projects rely on natural processes – for example, the expression of fusion proteins in E.coli and immune responses of small mammals – it’s hard to estimate the length of individual projects precisely and accurately. As a general rule, 4-6 months is a good starting point. At every stage of the project, our technical team will always inform you about the progress of your custom antibody production.

To receive a custom production quotation, please fill in the contact form below:

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