Synthetic peptide conjugation

The methods described are suitable for use with immunogens derived from a variety of sources, for example, bacterially expressed fusion proteins, baculovirus-expressed proteins or synthetic peptides conjugated to suitable carrier proteins (see Chapter 2). Antibody responses to peptide immunogens often differ from those where the immunogen is a larger macromolecule in that maximal antipeptide titres (which arise rapidly after two to three immunizations) do not always coincide with maximal titres against the intact... 

In a 10-mL syringe, with the open tip blocked with parafilm, add 5 mg of synthetic peptide conjugate and bring to a total volume of 5 mL with physiological saline. After this is mixed, add 5 mL of Complete Freund s adjuvant to the syringe... 

Wang TT, Fellows PF, Leighton TJ et al (2004) Induction of opsonic antibodies to the gamma-D-glutamic acid capsule of Bacillus anthracis by immunization with a synthetic peptide-carrier protein conjugate. FEMS Immunol Med Microbiol 40 231-237... 

Briand, J.P., Muller, S., and Van Regenmortel, M.H.V. (1985) Synthetic peptides as antigens pitfalls of conjugation methods./. Immunol. Meth. 78, 59-69. 

Boeckler C, et al. Immunogenicity of new heterobifunctional cross-linking reagents used in the conjugation of synthetic peptides to liposomes. J Immunol Meth 1996 191 1. 

Several synthetic methods are available for the preparation of Fc peptide conjugates, of which the active ester method is most compatible with biological environments.41 This method works under mild conditions with an isolable FcCO-active ester, in which reactive heterocyclic isolable esters are formed. These can be isolated or reacted in situ with suitable peptides to give the desired Fc-peptide conjugate. These active esters can be used as stoichiometric Fc delivery reagents, which make them suitable for automated solid-phase synthesis of Fc-peptide conjugates. 

Standard curve in competitive ELISA, 217-218 Streptavidin see avidin Stripping in immunoblotting, 234 Synthetic peptide, as antigen, 13-16 conjugation of, 19-23 synthesis of, 17-18... 

Key Words Synthetic peptide antibody antigen polyclonal immunogen carrier conjugation. 

Examination of presence of samples on array To verily that proteins, synthetic peptides, and carbohydrates are successfully printed, microarrays can be incubated with antibodies, receptors, or lectins known to react with the printed substance. The reaction is detected either by conjugating directly afluorochrome to the detector, or by a second-step staining procedure. 

Robey, F. A. and Fields, R. L. (1989) Automated synthesis of bromoacetyl-modi-fied peptides for preparation of synthetic peptide polymers, peptide-protein conjugates and cyclic peptides. Anal. Biochem. 177, 373-377. 

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