Premium
Development of a novel, quantitative protein microarray platform for the multiplexed serological analysis of autoantibodies to cancer‐testis antigens
Author(s) -
BeetonKempen Natasha,
Duarte Jessica,
Shoko Aubrey,
Serufuri JeanMichel,
John Thomas,
Cebon Jonathan,
Blackburn Jonathan
Publication year - 2014
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.28832
Subject(s) - antigen , autoantibody , protein microarray , cancer , immunology , microarray , cancer vaccine , cancer biomarkers , antibody , biology , monoclonal antibody , immune system , medicine , cancer research , immunotherapy , gene expression , gene , genetics
The cancer‐testis antigens are a group of unrelated proteins aberrantly expressed in various cancers in adult somatic tissues. This aberrant expression can trigger spontaneous immune responses, a phenomenon exploited for the development of disease markers and therapeutic vaccines. However, expression levels often vary amongst patients presenting the same cancer type, and these antigens are therefore unlikely to be individually viable as diagnostic or prognostic markers. Nevertheless, patterns of antigen expression may provide correlates of specific cancer types and disease progression. Herein, we describe the development of a novel, readily customizable cancer‐testis antigen microarray platform together with robust bioinformatics tools, with which to quantify anti‐cancer testis antigen autoantibody profiles in patient sera. By exploiting the high affinity between autoantibodies and tumor antigens, we achieved linearity of response and an autoantibody quantitation limit in the pg/mL range—equating to a million‐fold serum dilution. By using oriented attachment of folded, recombinant antigens and a polyethylene glycol microarray surface coating, we attained minimal non‐specific antibody binding. Unlike other proteomics methods, which typically use lower affinity interactions between monoclonal antibodies and tumor antigens for detection, the high sensitivity and specificity realized using our autoantibody‐based approach may facilitate the development of better cancer biomarkers, as well as potentially enabling pre‐symptomatic diagnosis. We illustrated the usage of our platform by monitoring the response of a melanoma patient cohort to an experimental therapeutic NY‐ESO‐1‐based cancer vaccine; inter alia , we found evidence of determinant spreading in individual patients, as well as differential CT antigen expression and epitope usage.