Peptides enzyme-linked immunosorbent assays

Kontrawelert, R, Francis, D.L., Brooks, RM., Ghosh, R. (1989) Application of an enzyme-linked immunosorbent-inhibition assay to quantitate the release of KS peptides into fluids of the rat subcutaneous air pouch model and the effects of chondroprotective drugs on the release process. Rheumatology International, 9 (2), 77-83. 

For cell surface antigens, the most frequently used assays are immunofluorescence or immunoenzymatic, with their numerous variations. For soluble protein or peptide antigens, the most common assays are immunoenzymatic, enzyme-linked immunosorbent assays (ELISA) or Western blot tests. 

Prince HE. Evaluation of the INOVA diagnostics enzyme-linked immunosorbent assay kits for measuring serum immunoglobulin G (IgG) and IgA to deamidated gliadin peptides. Clin Vaccine Immunol 2006 13 150-151. 

Accurate measurement of proinsulin has been difhcult for several reasons the blood concentrations are low antibody production is difficult most antisera cross-react with insulin and C-peptide, which are present in much higher concentrations the assays measure intermediate cleavage forms of proinsulin and reference preparations of pure proinsulin are not readily available. However, a more sensitive nonequiUb-rium RIA method for measuring proinsiilin was developed by adsorbing the initial antiserum with biosynthetic human C-peptide coupled to agarose to eliminate cross-reactivity with C-peptide.An enzyme-linked immunosorbent assay (ELISA) has been described that employs an antibody to C-peptide as the coating antibody and antiinsulin antibody for detection. The detection limit is 0.25 pmol/L. ... 

Gregorius, K., Theisen, M. (2001). In situ deprotection a method for covalent immobilization of peptides with well-defined orientation for use in solid phase immunoassays such as enzyme-linked immunosorbent assay. Anal. Biochem., 299, 84-91. 

Many types of assay are available to be used in HTS protocols to identify inhibitors of PPIs, but a competition assay, in which inhibition of complex formation is measured, is most common. Fluorescence polarization (FPA), fluorescence resonance energy transfer (FRET), enzyme-linked immunosorbent assays (ELISA), and other assay formats have been used. The interacting proteins can be used in their full-length forms though, more frequently only the interacting domains are employed, and if possible the excised interacting peptide is usually preferred. 

Cho, J-J., and Y-S. Lee. 1998. Enzyme linked immunosorbent assay for serum procollagen type III peptide in rats with hepatic fibrosis. Journal of Veterinary Medical Science 60 1213-1220. 

A second technique that accomplishes peptide synthesis on polyethylene rods was developed by Geyson et al.l95 A typical array of rods (called pins) consists of 96 pins (typically 4 mm in diameter and 40 mm in length) 194 arranged in 8 rows of 12 rods (see Figure 10.28b). The pins fit into the wells of plates (see Figure 10.28)194 that were developed for the enzyme-linked immunosorbent assay (ELISA). The polyethylene rods are functionalized with acrylic acid and NP-Fmoc-(3-alanyl-l,6-diaminohexane is attached as a spacer (see Figure 10.28a). The idea is to attached an amino acid to each of the pins, and place the solution containing the reactants into the wells. The pins are dipped into the wells and allowed to react. To deprotect the terminal... 

Kingan, T. (1989). A Competitive Enzyme-linked Immunosorbent Assay Applications in the Assay of Peptides, Steroids, and Cyclic Nucleotides, Biochem. 183 283-289. 

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