Novel type of protein chip for multiplex detection of autoantibodies

Abstract

Evaluation of: Akada J, Kamei S, Ito A et al. A new type of protein chip to detect hepatocellular carcinoma-related autoimmune antibodies in the sera of hepatitis C virus-positive patients. Proteome Sci. 11(1), 33 (2013).

Unlocking the proteome and delivering biomarkers to the clinic will be critical for early and improved diagnosis and prognosis. Conventional protein microarrays have evolved as a promising proteomic technology with great potential for protein expression profiling in health and disease. In this study, Akada et al. explore a new type of protein chip, interfaced with a dual-color fluorescence-based read-out, for screening of autoantibodies in serum. Uniquely, the recombinant antigens were microarray adapted by molecular design to contain a five-cysteine tag for immobilization and green fluorescent protein for detection (color 1). The engineered antigens were immobilized on in-house-designed maleimide-incorporated diamond-like carbon substrates and subsequently heat treated in a solution of denaturing and reducing agents before any specifically bound serum autoantibodies were detected (color 2). The authors used a 4-plex array targeting hepatocellular carcinoma-related autoantibodies in the sera of hepatitis C virus-positive patients as model system to demonstrate proof-of-concept.

Keywords::

• biomarker discovery
• denatured probes
• immobilization
• oligo-cysteine tag
• protein array
• protein chip

Financial & competing interests disclosure

The author is a co-founder of a small startup biotechnology company that uses antibody-based microarrays for detecting disease-associated biomarkers. The author was supported by grants from Greta and Johan Kock Foundation, the Swedish National Research Council (VR-NT and VR-M), SSF – the foundation of Strategic Research (Strategic Center for Translational Cancer Research – CREATE Health) and Vinnova. The author has no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Key issues

• • The paper under evaluation used proteins probes microarray adapted by molecular design.

• • The arrayed proteins were engineered with a C-terminal cysteine-tag for immobilization.

• • The arrayed proteins were engineered with an N-terminal GFP-tag for detection.

• • The arrayed proteins were on-chip denatured for improved array detection.

• • The novel protein chip was used to detect disease-associated serum autoantibodies.