Synthetic peptide as antigen

Muller (Eds), Synthetic Peptides as Antigens, Elsevier, Amsterdam, pp. 281 -317. 

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. 

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... 

The use of isotope-labelled synthetic peptides as IS was proposed for the absolute quantification of proteins in protein expression studies [113]. If necessary, these synthetic peptides can be covalently modified to be applied as IS in for instance phosphopeptide quantification. This approach was applied in the quantitative analysis of two glutathione 5 -transferase isoforms in human liver cytosol by LC-MS in SRM mode [114]. A series of pilot experiments were performed to select the most suitable IS peptides for four human plasma proteins (hemopexin, ocl antichymotrypsin, interleiddn-6, and tumor necrosis factor-oc) [115]. Rabbit polyclonal antibodies were raised against these selected peptides and immobilized on POROS supports. These lAC columns were applied to achieve a 120-fold emichment of the antigen peptide from digested human plasma proteins. The peptides and its IS were measured by LC-MS in SIM or SRM mode. The methods appears to be a tailor method for targeted protein analysis. 

Fig. 8.10 Titers of antibodies at day 50 induced by plant-derived CTB-2L21 recombinant protein. Balb/c mice were intraperitoneally immunized with leaf extract from CTB-2L21 transgenic plants. Animals were boosted at days 21 and 35. Each mouse received 20 pg of CTB-2L21 recombinant protein. Individual samples of mouse serum were titrated against 2L21 synthetic peptide,VP2 protein and a control peptide (amino acids 122-135 of hepatitis B virus surface antigen). Titers were expressed as the highest serum dilution to yield twice the absorbance mean of preimmune sera. M1-M6 mice 1 to 6 2L21 epitope from the VP2 protein of the canine parvovirus CTB cholera toxin B VP2 protein of the canine parvovirus that includes the 2L21 epitope.

Polyclonal antisera are usually prepared either from isolated antigens, such as proteins or from synthetic peptides derived from the protein sequence. The antisera derived from small synthetic peptides are regio-specific with respect to the cognate protein and resemble in some way the monoclonal antibodies, but they are much easier to prepare. 

---