Direct coating of poly(lys) or acetyl-thio-acetyl peptides to polystyrene: the effects in an enzyme-linked immunosorbent assay.
Direct adsorption of small peptides to polystyrene surfaces is often not satisfactory. Therefore, a simple and general coating procedure to improve the coating efficiency of small synthetic peptide antigens to polystyrene is described. In this study, the binding capacities of four small synthetic peptides N-terminally linked to various moieties during synthesis were compared to their parent counterparts in terms of the amount of peptide coat concentration required to achieve 50% of the maximum enzyme-linked immunosorbent assay signal. Elongation of a short epitope sequence by an N-terminal acetyl-thio-acetyl (Ata) group or a lysyl moiety resulted in an enormous reduction in peptide coat concentration for all tested peptides of net two to four orders of magnitude when corrected for chain elongation. The optimal length of the lysyl moiety depended on the length of the model peptide. Replacement of both extensions by analogues (i.e., Ata analogues and other basic amino acid residues in the case of the lysyl moiety) was possible without reducing their enhancing properties to a great extent. Additional experiments showed that a lysyl moiety consisting of a linear stretch of seven lysyl moiety consisting of a linear stretch of seven lysyl residues was more effective in comparison to a branched lysyl construct and could easily compete with the multiple antigen peptide approach.
Loomans
E E
EE
AKZO NOBEL, Chemistry Research Unit, Organon Teknika B. V., AB Boxtel, The Netherlands.
Petersen-van Ettekoven
A
A
Bloemers
H P
HP
Schielen
W J
WJ
D000107:Acetylation; D000327:Adsorption; D000595:Amino Acid Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006863:Hydrogen-Ion Concentration; D016014:Linear Models; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011107:Polylysine / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D000939:Epitopes; D010455:Peptides; D011137:Polystyrenes; D013438:Sulfhydryl Compounds; D011107:Polylysine