Polypeptide Semicarbazide Glass Slide Microarrays: Characterization and Comparison with Amine Slides in Serodetection Studies

Abstract

We have described in the accompanying article the preparation of peptide-protein semicarbazide microarrays and their use for the simultaneous serodetection of antibodies directed against different pathogens. Here, we present a comparative study between semicarbazide and amine glass slides in an immunofluorescent serodetection assay using HIV (Gp120, Gp41), HCV (mix-HCV, core, NS3, and NS4), and HBV (HBs) recombinant antigens. Amine and semicarbazide surfaces displayed the same sensitivity for antibodies detection just after printing. However, the reactivity of protein antigens changed rapidly upon aging on amine slides but not on semicarbazide slides. Peptide or protein semicarbazide microarrays were found to be remarkably stable for months. Additional data concerning the characterization of the semicarbazide surface (homogeneity of the slides, chemical stability, contact angle measurements, atomic force microscopy studies, reproducibility of serodetection results) are also presented and discussed.

Ambient-ageing processes in amine self-assembled monolayers on microarray slides as studied by ToF-SIMS with principal component analysis, XPS, and NEXAFS spectroscopy

“…Positive and negative ion spectra were internally calibrated using H + , H 2 + , CH 3 + , C 2 H 2 + , C 3 H 5 + and H − , C − , CH − , C 2 H − , and C 4 H − peaks, respectively. The total ion dose was less than 1.0×10 [12] ions/cm 2 [2] and well below the static limit of SIMS (∼1.0×10 [13] ions/cm 2 [2]). Effective charge compensation for samples on glass slides was obtained by using a pulsed electron flood gun.…”

“…Ageing influences technologically important parameters as morphology, wettability, and stability expressed as reactivity to, e.g., proteins [11][12][13]. Usually, users of amine slides store them for several days to months prior to use under rather different conditions.…”

Protein‐Detecting Microarrays: Current Accomplishments and Requirements

“…[56] The same group also constructed protein microarrays in which proteins were adsorbed onto the semicarbazide-modified surfaces, and they analyzed interactions with HIV (gp120, gp41), HCV (mix-HCV, core, NS3, and NS4), and HBV (HBs) recombinant antigens. [157] Meanwhile, immunoglobulin E (IgE) epitope mapping of food allergens is a prerequisite for engineering hypoallergenic immunotherapeutic agents and can provide important information regarding a patient's immune response. Schreffler et al developed an immunoassay based on a peptide microarray to map peanut epitopes.…”

Development of a fluorescent and colorimetric detection methods-based protein microarray for serodiagnosis of TORCH infections

“…To minimize the disadvantages, various chemical surfaces on glass slides were tested with a number of coupling chemistries, such as amine (Duburcq et al, 2004a), aldehyde (Zammatteo et al, 2000), poly-l-lysine (Haab et al, 2001), epoxy , agarose (Afanassiev et al, 2000), polyacrylamide (Angenendt et al, 2002) and nitrocellulose (Qiu et al, 2004). It is reported that polyacrylamide slides showed a sixfold signal-to-noise ratio higher than poly-l-lysine slides coated with an additional photo-reactive crosslinker (HSAB slides) when applied to protein profiling of prostate cancer (Miller et al, 2003).…”