Multiple Peptide Co-immunization

Select your desired Custom Multiple Peptide Co-immunization Polyclonal Antibody Services using Bio-Synthesis all inclusive production packages that are scientifically proven, species-specific schedules in rabbits. Should you not find a package that suits your needs, you have the option to developing a customized host species and immunization schedule to meet your protein detection needs.
Request a quote or contact us for your Multiple Peptide Co-immunization packages.

Multiple-Peptide Co-immunization

While multiple-peptide co-immunization is designed to produce a good polyclonal antibody that can capture the protein from multiple epitopes is produced. However, in practice all antigens are not equal in immunizing potential. Very often the immunodominant peptide will produce the overwhelming majority of antibodies, and little ground is gained for the added expense.

Bio-Synthesis will synthesize, purify and conjugate each peptide separately. Individual KLH conjugates are then mixed in equimolar ratios and used as immunogenic. Individual antisera are titered by ELISA against each peptide separately. We offer four peptide cocktail custom antibody production packages, each of which provides a minimum of four antibodies against your protein of interest. Should you not find a package that suits your needs, we will design one for you.

Consult with our peptide and antibody production teams to help with the best decision regarding multiple peptide co-immunization options.

Strategies for Cocktail-peptide Co-immunization

Two strategies for multiple-peptide co-immunization

Homogeneous peptide conjugation and immunization - This appears to be the more preferred strategy in multiple-peptide co-immunization, in which several peptide sequences represent different epitopes of the same protein is conjugated into a protein carrier separately and each conjugate is then purified and characterized. These different peptide-protein conjugates are mixed together and formulated for immunization. Since each carrier protein molecular is conjugated only to one type of peptides, this may enable distinct recognition and T-cell responses to each peptide antigen. The antibodies can be isolated as a group responding to all peptide epitopes, or as individual antibodies responding to a specific peptide epitope.

Heterogenous peptide conjugation and immunization - This is where several different peptides representing different epitopes of the same protein are mixed together before conjugation. The mixture of peptides is then conjugated as one batch into a carrier protein. While this strategy has an advantage of using a single conjugation step for all peptides in the mixture, the obvious assumption here is that each peptide sequence will conjugate with equal efficiency to the carrier protein. However, this will not be easy to prove because it will be difficult to characterize each peptide-protein conjugate. It is also uncertain whether or not each peptide epitope will be equally recognized by the immune system and elicit antibodies to the same extent. In this approach, the antibodies can also be isolated as a group responding to all peptide epitopes, or as individual antibodies responding to a specific peptide epitope.

Several other types of anti-peptide polyclonal antibody services also offered in our laboratory

We offer four Multiple Peptide Co-immunization Polyclonal Antibody Service Packages. User-design production protocol also available and can also be modified from existing package.

Serum Production Specification:

Continuation of project past our standard immunization schedule is offered on a month to month basis. The standard extension protocol includes two boosters, one serum collection, and maintenance for one month. Terminal bleeds may be obtained at any time yielding approximately ~50 ml/rabbit of additional serum taken upon customer request.

Program Development of Rabbit Polyclonal Antibody: 70-90 Days

Procedure Protocol Days * Description Phase I - Antigen design, synthesis and preparation Antigen Preparation ~5 - 10 days Peptide synthesis and purification (if required) and antigen conjugation Phase II - Immunization Schedule Control sera collection Initial Inoculation Day 0 NZQ female rabbits and Control serum (~5 ml serum/rabbit) Primary Immunization with 500 µg peptide conjugate or 250 µg protein per host with FCA in 10 SQ sites 1st Booster Day 14 Boost with 250 µg peptide conjugate or 125 µg protein per host with FIC in 4 SQ sites 2nd Booster Day 28 Boost with 250 µg peptide conjugate or 125 µg protein per host with FIC in 4 SQ sites 1st Serum Collection 3rd Booster Day 42 Week 6 Collection Serum ~25 ml/rabbit Boost with 250 µg peptide conjugate or 125 µg protein per host with FIC in 4 SQ sites 2nd Serum Collection 4th Booster ELISA Titer Assay Day 56- 63 Week 8 Collection Serum ~25 ml/rabbit ELISA assay of control and week 8 collection serum ction serum 1st Shipment Project Completion Verification Day 63-64 Shipping of control, week 6 and 8 collection serum Verify disposition of rabbits, extension or termination 3rd Serum Collection Day 70 Week 10 Collection Serum ~25 ml/rabbit Final Shipment l Shipment Day 75 Shipping of week 10 collection serum Instruction Due Date on Due Date Day 84 Termination of animals if instruction not received ot received

Program Development of Rabbit Polyclonal Antibody: 90-120 Days

Procedure Protocol Days * Description Phase I - Antigen design, synthesis and preparation Antigen Preparation ~5 - 10 days Peptide synthesis and purification (if required) and antigen conjugation Phase II - Immunization Schedule Control serum collection Initial Inoculation Day 0 NZQ female rabbits and Control serum (~5 ml serum/rabbit) Initial Inoculation with 500 µg peptide conjugate or 250 µg protein per host with FCA in SQ 4 sites 1st Booster Day 14 Boost with 500 µg peptide conjugate or 125 µg protein per host with FIC in SQ 4 sites 2nd Booster Day 28 Boost with 250 µg peptide conjugate or 125 µg protein per host with FIC in 4 SQ sites Test Serum Collection 3rd Booster Day 42 Week 6 Test serum 2-5 ml/rabbit Boost with 250 µg peptide conjugate or 125 µg protein per host with FIA in SQ 4 sites 1st Serum Collection 4th Booster Day 56-65 Week 8 Collection serum ~25 ml/rabbit Boost with 250 µg peptide conjugate or 125 µg protein per host with FIC in 4 SQ sites 2nd Serum Collection 5th Booster ELISA Titer Assay Day 70-75 Week 10 Collection Serum ~25 ml/rabbit Boost with 250 µg peptide conjugate or 125 µg protein per host with FIC in 4 SQ sites ELISA assay of control, week 6 test serum and 10 collection serum 1st Shipment Project Completion Verification Day 77 Shipping of control, 6, 8, 10 collection serum and ELISA report Verify disposition of rabbits, extension or termination 3rd Serum Collection 2nd Shipment Day 84 Week 12 Collection Serum ~25 ml/rabbit 2nd Shipment of week 12 collection serum Instruction Due Date  Day 98 Termination of animals if instruction not received Phase III - Post Production Sample Processing Serum Processing 2-3 weeks affinity column preparation, purification and/or labeling ELISA and/or SDS Analysis 1 week Analysis of purified sample Final Shipment Day 108 Shipping of purified sample, unpurified week 12 serum and documents

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