Multiplex Quantification of Plasma Proteins

Soluble proteins were quantified in EDTA-anticoagulated plasma using the Luminex multiplex platform (Luminex) with custom-developed reagents (R&D Systems), as described in detail (Guiducci et al., 2006 (link)) or single-plex ELISA (R&D Systems). Analytes quantified using the Luminex multiplex platform were read on the MAGPIX instrument and raw data were analyzed using the xPONENT software. Analytes quantified using single-plex ELISA were read using optical density. Values outside the lower limit of quantification were assigned a value of one-third of the lower limit of the standard curve for analytes quantified by Luminex and half of the lower limit of the standard curve for analytes quantified by ELISA. CXCL4 concentrations were determined using CXCL4/PF4 Quantikine (from R&D Systems) ELISA kit.

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Çakan E., Ah Kioon M.D., Garcia-Carmona Y., Glauzy S., Oliver D., Yamakawa N., Vega Loza A., Du Y., Schickel J.N., Boeckers J.M., Yang C., Baldo A., Ivashkiv L.B., Young R.M., Staudt L.M., Moody K.L., Nündel K., Marshak-Rothstein A., van der Made C.I., Hoischen A., Hayward A., Rossato M., Radstake T.R., Cunningham-Rundles C., Ryu C., Herzog E.L., Barrat F.J, & Meffre E. (2023). TLR9 ligand sequestration by chemokine CXCL4 negatively affects central B cell tolerance. The Journal of Experimental Medicine, 220(12), e20230944.

Publication 2023

Luminex multiplex platform custom-developed reagents single-plex ELISA MAGPIX instrument

Corresponding Organization : Cornell University

Other organizations : Bridge University, Icahn School of Medicine at Mount Sinai, National Cancer Institute, National Institutes of Health, University of Massachusetts Chan Medical School, Radboud University Nijmegen, Radboud University Medical Center, Brown University, Providence College, University of Verona, University Medical Center Utrecht, Utrecht University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Soluble proteins quantified in EDTA-anticoagulated plasma using the Luminex multiplex platform
Soluble proteins quantified using single-plex ELISA
CXCL4 concentrations determined using CXCL4/PF4 Quantikine ELISA kit

control variables

EDTA-anticoagulated plasma
Luminex multiplex platform with custom-developed reagents (R&D Systems)
Single-plex ELISA (R&D Systems)
CXCL4/PF4 Quantikine ELISA kit (R&D Systems)

controls

Positive control: Not specified
Negative control: Not specified

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