SACRI antibody services (SAS) now offers a complete anti-peptide antibody production package which begins with synthesis of your antigenic peptide.  All our custom anti-peptide projects include a peptide design consultation.  Our valuable experience producing anti-peptide antibodies provides clients with the maximum likelihood of successful results.  Our site has many online resources that can help you design the best synthetic peptide for use as an immunogen.

SAS consults directly with clients at each critical stage in the production of anti-peptide antibodies.  Our protocol involves three important stages.

Stage 1: Peptide selection and design

SAS custom anti-peptide antibody packages include a free consultation with our peptide design experts, however, we suggest you first review our peptide design page for helpful tips and suggestions.  After our clients submit a potential epitope sequence, our immunochemists and protein chemists will work together to analyze the sequence and ensure the peptide can be successfully synthesized and will prove immunogenic.  Our SAS staff will then consult the client as to the suitability of the peptide and either continue to the next stage or revisit the design process until a consensus is reached as to the best potential sequence.

Stage 2: Peptide synthesis and quality assurance

The peptide is produced in our Peptide Synthesis Laboratory by a group of skilled and dedicated protein chemists using FMOC solid phase chemistry.  Each peptide will have >70% purity, is tested using mass spectrometry and HPLC to ensure the finest sample for immunization.  Peptides can be made with custom modifications, MAP (multiple antigenic peptide) linked, or conjugated to a number of carrier molecules (e.g. KLH, BSA, etc.).  After the quality of the synthesized peptide is assured, a sample mass spectrometry and HPLC trace will be provided to our client.  If all the modifications and conjugations are satisfactory, our immunochemists will take the sample and continue with antibody production.

Stage 3: Animal immunization and antibody production

SAS utilizes a standard 70 day protocol with one or more animals.  Polyclonal antibodies are produced by immunizing host animals against the conjugated peptide antigen.  Animals are given 4 subsequent boost injections to increase the success rate of the protocol.  Blood samples are collected at regular intervals and titre levels are tested using by ELISA. When the antibody titre reaches a maximal level a cardiac puncture is preformed.  All the serum is collected for a final purification procedure.

Animals can be kept for longer periods of time than the basic injection schedule at a fee. Contact us to design a customized protocol for your research needs.

SAS also provides a protein purification service to increase the concentration of antibodies in the delivered serum.

Quality control measures

Several quality control measures are available to ensure clients are supplied with only the highest quality antibodies including:

ELISA, SDS/PAGE, isoelectric focusing, HPLC, Western Blot

Additional functional tests can be performed upon the clients' request.

Pricing

SAS provides only custom anti-peptide antibody packages and pricing will vary depending upon the exact peptide and antibody requirements of our clients.  Stages 1 and 2 typically require 10 days and are estimated to cost $400-$600 depending upon your requirements.

Our standard polyclonal development procedure requires 2 months to complete.  Exact costs will depend upon the specific requirements of each client, including choice of host animal, the number of animals immunized and other variables.

Order custom anti-peptide antibodies from SAS

Frequently asked questions - custom anti-peptide antibodies

Q: How do I select an appropriate peptide sequence?

A: Although the design of peptides for use as antigens is not exact, several hints can help maximize the likelihood of success in producing antibodies.  It is important to select sequences which are found on the surface of the native protein, and contain residues that are hydrophilic and have flexible side-chains.  A BLASTp search should be performed and a sequence with minimal homology selected to reduce the chance of non-specific antibody binding.  Also, peptide sequence length, hydrophobicity of residues, and specific sequence issues are critical.  See our peptide design page for more detailed suggestions.

Q: What are the advantages to using synthetic peptides as antigens?

A: One of the main advantages to using synthetic peptides for antibody production is that specific epitope can be targeted.  Peptides can be designed and modified to contain precisely what is required for your experiment.  Synthetic peptides are also relatively inexpensive to produce and can be obtained in a highly purified form.

Q: What peptide purity do I require?

A: For use as an immunogen, peptide purity (expressed as the % of correct amino acids in the peptide) should be at least 70%.

Q: Will my peptide be difficult to synthesize?

A: Several factors will influence the production and purification of a peptide including its length and amino acid composition, hydrophobicity and stretches of the same residue.  Our experienced protein chemists will help review your sequence to determine whether the peptide can be synthesized at sufficient purity for use as an immunogen.

Q: How long will it take to make an anti-peptide antibody?

A: We provide an extremely fast turn around time from the receipt of a clients' order to delivery of the antibody.  Peptide design and synthesis is generally complete after 10 days, and our standard polyclonal antibody protocol requires 70 days, so our clients should receive their custom antibody after 2.5 months.

Q: Which carrier molecules are available, and which should I use?

A: Conjugation of a peptide to a carrier molecule is critical since peptide molecules alone often fail to initiate an immune response.  Carrier molecules contain multiple epitopes that can stimulate T-helper cells, which lead to a B-cell response.  Keyhole-limpet hemocyanin (KLH) and bovine serum albumin (BSA) are the most common protein carriers, and KLH is preferred since it is more antigenic in the majority of animals.  Also, BSA is often used as a blocking reagent in assays, thus an anti-peptide antibody raised against a BSA conjugated peptide will show some specificity towards the buffer reagent.  This can result in a false positive signal.

Q: Should I use a MAP (multiple antigenic peptide)?

A: MAP is simply an alternative to KLH conjugation.  It is recommended only for short peptides located either internally within a protein or at the N-terminus.  KLH conjugation is recommended for a peptide at any location in a protein and when there is a cysteine residue within the peptide sequence.

Q: What are the yields of antisera for type of bleed in the different host animals?

A:

Order custom anti-peptide antibodies from SAS

Send mail to Sampson Law with questions or comments about this web site.
Last modified: 09/15/05 00:00:00