A quantitative assay to measure the interaction between immunogenic peptides and purified class I major histocompatibility complex molecules

✍ Scribed by Anna Catharina Olsen; Lars ØStergaard Pedersen; Anette Stryhn Hansen; Søren Buus; Mogens Holst Nissen; Marianne Olsen; Paul Robert Hansen; Arne Holm

• Publisher: John Wiley and Sons
• Year: 1994
• Tongue: English
• Weight: 867 KB
• Volume: 24
• Category: Article
• ISSN: 0014-2980
• DOI: 10.1002/eji.1830240218

✦ Synopsis

A direct and sensitive biochemical assay to measure the interaction in solution between peptides and affinity-purified major histocompatibility complex (MHC) class I molecules has been generated. Specific binding reflecting the known class I restriction of cytotoxicT cell responses was obtained. Adding an excess of P*-microglobulin (pzm) significantly increased the rate of peptide association, but it did not affect the rate of dissociation. Binding was complicated by a rapid and apparently irreversible loss of functional MHC class I at 37°C which might limit the life span of empty MHC class I thereby preventing the inadvertent exchange of peptides at the target cell surface. All class I molecules tested bound peptides of the canonical octa-to nona-meric length. However, one class I molecule, Kk, also bound peptides, which were much longer suggesting that the preference of class I molecules for short epitopes is not absolute and may be caused by factors other than the peptide-MHC class I binding event itself.