Passive adsorption of immunologically active and inactive synthetic peptides to polystyrene is influenced by the proportion of non-polar residues in the peptide.
A well-known drawback in the use of synthetic peptides as solid-phase antigens in immunoassays is that positive controls confirming the presence of the peptide on the solid phase are not always present. We therefore evaluated the applicability of a recently described enzyme immunoassay (EIA) method by which the presence of peptides is detected by biotinylation (BioEIA) of alpha- and/or epsilon-amino groups after passive adsorption. This approach allows the rapid screening of a large number of proteins and peptides in respect to passive adsorption to plastic surfaces. When using irradiated polystyrene microplates we found that 240 (94%) of 256 synthetic peptides, covering 85% of the complete hepatitis C virus (HCV) sequence, passively adsorbed to polystyrene. When comparing the results from the BioEIA to the peptide reactivity of human sera it was obvious that the absence of serum reactivities was not due to lack of peptide adsorption to the plates. Using 192 peptides the relation between the signal-to-cutoff ratio (S/CO) in the BioEIA and the amino acid content of the individual peptides was further analyzed. The S/CO ratio was related to the number of epsilon NH2 groups (Lys residues) present in the peptide (P < 0.001, Kruskal-Wallis). We separately related the amino acid content of 68 peptides with Lys and 124 peptides lacking Lys to the S/CO ratio in the BioEIA. In both cases it was found that an increasing amount of nonpolar residues such as Ala, Phe, Ile, Met, and Val (P < 0.05, respectively) in the peptides was related to a lower S/CO ratio in the BioEIA.(ABSTRACT TRUNCATED AT 250 WORDS)
Sällberg
M
M
Department of Immunology, Microbiology, Pathology, and Infectious Diseases, Karolinska Institutet, Huddinge, Sweden.
Blixt
M
M
Zhang
Z X
ZX
Ekstrand
J
J
D000327:Adsorption; D000596:Amino Acids / Q000737:chemistry* / Q000502:physiology; D000941:Antigens / Q000737:chemistry*; D016174:Hepacivirus / Q000737:chemistry / Q000276:immunology; D007124:Immunoenzyme Techniques; D007163:Immunosorbent Techniques; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011137:Polystyrenes / Q000737:chemistry*; D011485:Protein Binding / Q000276:immunology*; D014758:Viral Core Proteins / Q000737:chemistry / Q000276:immunology
D000596:Amino Acids; D000941:Antigens; D010455:Peptides; D011137:Polystyrenes; D014758:Viral Core Proteins