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Bio plex pro human isotyping assay

Manufactured by Bio-Rad
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The Bio-Plex Pro Human Isotyping Assay is a multiplex assay designed to simultaneously quantify up to 11 human immunoglobulin isotypes from a single sample. The assay utilizes magnetic beads coated with specific capture antibodies for each isotype, allowing for the measurement of IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgM, IgD, and IgE in a single well.

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Lab products found in correlation

Raybio human egf elisa kit, by RayBiotech (1 mentions) Bio plex 200 system instrument, by Bio-Rad (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Cpg oligodeoxynucleotide odn 2006, by InvivoGen (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by GE Healthcare (1 mentions) Bio plex 3d suspension array system, by Bio-Rad (1 mentions) April, by Thermo Fisher Scientific (1 mentions)

6 protocols using bio plex pro human isotyping assay

1

Serum Antibody Isotyping Assay

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Antibody isotypes IgG1, IgG2, IgG3, IgG4, IgA and IgM in serum were assessed in singlicate using the Bio‐Plex Pro™ Human Isotyping Assay (Bio‐Rad Corporation) according to manufacturer's instructions and analysed on the Bio‐Plex 200 system.
de Silva H.D., Ffrench R.A., Korem M., Orlowski E., Curtis D.J., Spencer A., Avery S., Patil S, & Morrissey C.O. (2018). Contemporary analysis of functional immune recovery to opportunistic and vaccine‐preventable infections after allogeneic haemopoietic stem cell transplantation. Clinical & Translational Immunology, 7(10), e1040.
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2

Biomarker Profiling of Human Milk

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Concentrations of Igs (IgA, IgG, and IgM), transforming growth factor-beta 2 (TGF-β2) and hormones (adiponectin, ghrelin, and leptin) were determined in duplicate using a Bioplex 200 system instrument (Bio-Rad, Hercules, CA, USA) and the Bio-Plex Pro Human Isotyping Assay, Bio-Plex Pro Human TGF-β Assay and Bio-Plex Pro Human Diabetes Assays kits (Bio-Rad). The concentration of EGF was measured using the RayBio® Human EGF ELISA kit (RayBiotech, Norcross, GA). Every assay was performed according to manufacturer's instructions. Standard curves were performed for each analyte. The analytes were assayed in, at least, three batches for each HTST treatment.
The inter-assay coefficients of variation were below manufacturers' instructions for all the immune markers, and the lower limit of quantification (LLOQ) for every analyte in human milk were: 0.21 μg/L for IgA, 0.78 μg/L for IgM, 2.19 μg/L for IgG, 1.57 ng/L for TGF-β2, 0.03 ng/L for EGF, 23.10 ng/L for adiponectin, 7.40 ng/L for ghrelin, and 11.45 ng/L for leptin.
Escuder-Vieco D., Espinosa-Martos I., Rodríguez J.M., Fernández L, & Pallás-Alonso C.R. (2018). Effect of HTST and Holder Pasteurization on the Concentration of Immunoglobulins, Growth Factors, and Hormones in Donor Human Milk. Frontiers in Immunology, 9, 2222.
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3

Profiling B Cell Immunoglobulin Isotypes

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Immunoglobulin heavy-chain isotypes of B cell clones were determined by analysis of cell culture supernatants of B cell clones using the Bio-Rad Bio-Plex Pro Human Isotyping Assay (Bio-Rad, Hercules, CA).
van de Veen W., Globinska A., Jansen K., Straumann A., Kubo T., Verschoor D., Wirz O.F., Castro-Giner F., Tan G., Rückert B., Ochsner U., Herrmann M., Stanić B., van Splunter M., Huntjens D., Wallimann A., Fonseca Guevara R.J., Spits H., Ignatova D., Chang Y.T., Fassnacht C., Guenova E., Flatz L., Akdis C.A, & Akdis M. (2020). A novel proangiogenic B cell subset is increased in cancer and chronic inflammation. Science Advances, 6(20), eaaz3559.
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4

Activation and Analysis of Human B Cells

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CD19+ B cells were isolated form PBMC and resuspended in Roswell Park Memorial Institute 1640 medium, supplemented with 10% heat-inactivated fetal bovine serum (GE Healthcare Sciences, Salt Lake City, UT, USA), 1.5% HEPES (Invitrogen), and 1% penicillin/streptomycin (Invitrogen). The 1 × 105 B cells were activated with 0.5 μg/mL BAFF (ProSpec, Vineland, NJ, USA) and/or 1 μg/mL CpG oligodeoxynucleotide (ODN) 2006 (InvivoGen, San Diego, CA, USA). After 48 h, the ASC differentiations were analyzed by flow cytometry, and the Ig levels in the undiluted culture supernatants were measured using Bio-Plex Pro Human Isotyping Assays (Bio-Rad).
Doi H., Hayashi E., Arai J., Tojo M., Morikawa K., Eguchi J., Ito T., Kanto T., Kaplan D.E, & Yoshida H. (2018). Enhanced B-cell differentiation driven by advanced cirrhosis resulting in hyperglobulinemia. Journal of gastroenterology and hepatology.
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5

Assessing Ig Isotypes and BAFF/APRIL

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Sera from HD and patients were freshly isolated and cryopreserved prior to analyses. Ig isotypes were measured with Bio-Plex Pro Human Isotyping Assays (Bio-Rad) on BioPlex 3D Suspension Array System. Enzyme-linked immunosorbent assay for BAFF (R&D Systems, Minneapolis, MN, USA) and A proliferation-inducing ligand (APRIL) (eBioscience, San Diego, CA, USA) were performed as per manufacturer’s instructions.
Doi H., Hayashi E., Arai J., Tojo M., Morikawa K., Eguchi J., Ito T., Kanto T., Kaplan D.E, & Yoshida H. (2018). Enhanced B-cell differentiation driven by advanced cirrhosis resulting in hyperglobulinemia. Journal of gastroenterology and hepatology.
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6

Quantifying Immunoglobulin Isotypes

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All maternal plasma, infant plasma, and BM supernatant was tested using the Bio-Rad Bio-Plex Pro Human Isotyping Assays (171A3100M) to quantify IgG1, IgG2, IgG3, IgG4, IgA, and IgM per manufacturer’s protocol. Samples were tested at a 1:40,000 dilution, and data were acquired on the MAGPIX through access from the Boston University Analytical Core. Total IgG was determined from summing quantities of IgG1, IgG2, IgG3, and IgG4.
Thomas A.S., Coote C., Moreau Y., Isaac J.E., Ewing A.C., Kourtis A.P, & Sagar M. (2022). Antibody-dependent cellular cytotoxicity responses and susceptibility influence HIV-1 mother-to-child transmission. JCI Insight, 7(9), e159435.
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