Exploration of high-density protein microarrays for antibody validation and autoimmunity profiling
2016 (English)In: New Biotechnology, ISSN 1871-6784, E-ISSN 1876-4347, Vol. 33, no 5, 582-592 p.Article in journal (Refereed) PublishedText
High-density protein microarrays of recombinant human protein fragments, representing 12,412 unique Ensembl Gene IDs, have here been produced and explored. These protein microarrays were used to analyse antibody off-target interactions, as well as for profiling the human autoantibody repertoire in plasma against the antigens represented by the protein fragments. Affinity-purified polyclonal antibodies produced within the Human Protein Atlas (HPA) were analysed on microarrays of three different sizes, ranging from 384 antigens to 21,120 antigens, for evaluation of the antibody validation criteria in the HPA. Plasma samples from secondary progressive multiple sclerosis patients were also screened in order to explore the feasibility of these arrays for broad-scale profiling of autoantibody reactivity. Furthermore, analysis on these near proteome-wide microarrays was complemented with analysis on HuProt (TM) Human Proteome protein microarrays. The HPA recombinant protein microarray with 21,120 antigens and the HuProt (TM) Human Proteome protein microarray are currently the largest protein microarray platforms available to date. The results on these arrays show that the Human Protein Atlas antibodies have few off-target interactions if the antibody validation criteria are kept stringent and demonstrate that the HPA-produced high-density recombinant protein fragment microarrays allow for a high-throughput analysis of plasma for identification of possible autoantibody targets in the context of various autoimmune conditions.
Place, publisher, year, edition, pages
Elsevier, 2016. Vol. 33, no 5, 582-592 p.
Antibodies, Antigens, Bioassay, Biochips, Microarrays, Proteins, Autoimmune conditions, Different sizes, High-throughput analysis, Polyclonal antibody, Protein fragments, Protein microarray, Secondary progressive multiple sclerosis, Validation criteria
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-189649DOI: 10.1016/j.nbt.2015.09.002ISI: 000378026000012PubMedID: 26417875OAI: oai:DiVA.org:kth-189649DiVA: diva2:949252
7th Alpbach Workshop on Affinity Proteomics, MAR 09-11, 2015, Alpbach, Austria
FunderScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 201607182016-07-182016-07-112016-07-18Bibliographically approved