Abstract
We used the microarray technology to develop chips containing a comprehensive set of proteins and peptides covering the proteome of HIV-1 clade C, which is the HIV-1 subtype that causes the majority of infections worldwide. We demonstrate that the HIV microarray allows simultaneous, sensitive and specific detection of antibody responses for the major immunoglobulin classes (IgG, IgA, IgM, IgE) and subclasses (IgG1-4) with minute amounts of serum samples towards a large number of HIV antigens and peptides. Furthermore, we show that the HIV chip can be used for the monitoring of antibody responses during the course of the disease and during treatment. The HIV microarray should be useful to study antibody responses to multiple HIV antigens and epitopes in HIV-infected patients to explore pathomechanisms of the disease, for diagnosis and for monitoring of treatment and of vaccine trials.
Highlights
We used the microarray technology to develop chips containing a comprehensive set of proteins and peptides covering the proteome of HIV-1 clade C, which is the HIV-1 subtype that causes the majority of infections worldwide
We demonstrate that the HIV microarray allowed the measurement of isotypes and IgG subclasses against a comprehensive set of proteins and peptides covering the clade C proteome
Design of a chip containing a comprehensive set of micro-arrayed peptides and proteins of the HIV-1 clade C proteome
Summary
We used the microarray technology to develop chips containing a comprehensive set of proteins and peptides covering the proteome of HIV-1 clade C, which is the HIV-1 subtype that causes the majority of infections worldwide. The aim of this study was the development of an HIV microarray containing a large panel of HIV proteins and peptides for the mapping and characterization of HIV-specific antibody responses towards multiple viral antigens and epitopes with minimal amounts of sample and short assay-duration. For this purpose, we employed the microarray-chip technology which we originally had developed for the diagnosis of allergy (i.e., Immuno solid-phase allergen chip, ISAC).[10] We prepared a set of HIV proteins and peptides derived from HIV-1 clade C, because this is the HIV-1 subtype that causes the majority of infections worldwide (48%). We demonstrate that the HIV microarray allowed the measurement of isotypes and IgG subclasses against a comprehensive set of proteins and peptides covering the clade C proteome
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