Properties of cryptic epitopes and their corresponding antibodies as indicated by the study of human and ovine growth hormones.
Antibodies (Ab) directed to hidden antigenic determinants (cryptotopes) are undesirable because they are not neutralizing. Additionally, we have previously demonstrated a close association between the extent of Ab to cryptic determinants and the expression of autoantibodies (autoAb) under some experimental conditions. Thus, the first objective of this work was to establish the physicochemical characteristics of Ab to cryptotopes and the second one was to examine the structural features of cryptic epitopes themselves. Using human and ovine growth hormones (hGH and oGH) as antigenic models and competition ELISA under different conditions of temperature, pH or ionic strength, we did not find any difference between the binding properties of anti-cryptic epitope antibodies (Ab) and anti-native epitope Ab. Then, using synthetic peptides and tryptic digests and direct and competition ELISAs we studied the structures of cryptic hGH and oGH epitopes. Isolated peptides either in solution or adsorbed on microplates failed to react. Partially digested hGH was recognized only when insolubilized on microplates, and anti-oGH Ab only reacted with a large fragment of the hormone either in solution or insolubilized. These results indicate that, at least in the case of hGH and oGH, cryptic epitopes are not simple linear sequences, as commonly referred without any evidence, but new exposed conformational structures different from those found in the native antigen.
Loureiro
M E
ME
Facultad de Farmacia y Bioquímica, Instituto de Química y Fisicoquímica Biológicas (UBA-CONICET), Buenos Aires, Argentina.
Marino
V J
VJ
Mathieu
P A
PA
Duhalde
M
M
Roguin
L P
LP
Peña
C
C
Retegui
L A
LA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000276:immunology*; D005260:Female; D013006:Growth Hormone / Q000276:immunology*; D019382:Human Growth Hormone / Q000276:immunology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008807:Mice, Inbred BALB C; D008808:Mice, Inbred CBA; D009994:Osmolar Concentration; D010455:Peptides / Q000737:chemistry; D012756:Sheep; D013696:Temperature; D014357:Trypsin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010455:Peptides; D019382:Human Growth Hormone; D013006:Growth Hormone; D014357:Trypsin