Human plasma

Manufactured by Complement Technology

Human plasma is a component of blood that is separated from the cellular elements. It is a clear, straw-colored liquid that contains a variety of proteins, electrolytes, and other substances. The core function of human plasma is to provide a medium for the transport of blood cells, nutrients, and other essential substances throughout the body.

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Lab products found in correlation

Fitc conjugated anti mouse igg, by Merck Group (2 mentions) Anti fh mab, by Quidel (2 mentions) Facscalibur, by BD (1 mentions)

2 protocols using human plasma

Measuring Bacterial Factor H Binding

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Factor H (FH) binding to bacteria was performed using flow cytometry as described previously (44 (link)). Briefly, N. gonorrhoeae F62 ΔlgtD harvested from chocolate agar plates was grown in liquid media that contained the specified concentration of the CMP-NulO. Bacteria were then washed with Hanks Balanced Salt Solution (HBSS) containing 1 mM CaCl₂ and 1 mM MgCl₂ (HBSS⁺⁺) and incubated with 20 µg/ml of FH purified from human plasma (Complement Technology, Inc.). Bound FH was detected using an anti-FH mAb (Quidel; clone 90X), followed by FITC conjugated anti-mouse IgG (Sigma); both Abs had similar performance characteristics. All reaction mixtures were carried out in HBSS⁺⁺/1% (w/v) BSA in a final volume of 50 µl.

Gulati S., Schoenhofen I.C., Lindhout-Djukic T., Schur M.J., Landig C., Saha S., Deng L., Lewis L.A., Zheng B., Varki A, & Ram S. (2020). Therapeutic CMP-nonulosonates against multidrug-resistant Neisseria gonorrhoeae. Journal of immunology (Baltimore, Md. : 1950), 204(12), 3283-3295.

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Bacterial Binding of Factor H Assay

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Factor H (FH) binding to bacteria was performed using flow cytometry as described previously [16 (link)]. Briefly, bacteria (Ng F62 ΔlgtD) were harvested from chocolate agar plates and grown in liquid media that contained the specified concentration of the CMP-NulO as described above. Bacteria were then washed with Hanks Balanced Salt Solution (HBSS) containing 1mM Ca²⁺ and 1 mM Mg²⁺ (HBSS⁺⁺) and incubated with FH purified from human plasma (Complement Technology, Inc.; concentration specified for each experiment). Bound FH was detected using either anti-FH mAb (Quidel, catalog no. A254 (mAb 90X)) or affinity-isolated polyclonal goat anti-human FH, followed by FITC conjugated anti-mouse IgG or anti-goat IgG, respectively (Sigma); both Abs had similar performance characteristics. All reaction mixtures were carried out in HBSS⁺⁺/1% BSA in a final volume of 50 μl. Flow cytometry was performed using a FACSCalibur instrument (Becton Dickinson) and data were analyzed using FlowJo (version 7.2.5; Tree Star, Inc.).

Gulati S., Schoenhofen I.C., Whitfield D.M., Cox A.D., Li J., St. Michael F., Vinogradov E.V., Stupak J., Zheng B., Ohnishi M., Unemo M., Lewis L.A., Taylor R.E., Landig C.S., Diaz S., Reed G.W., Varki A., Rice P.A, & Ram S. (2015). Utilizing CMP-Sialic Acid Analogs to Unravel Neisseria gonorrhoeae Lipooligosaccharide-Mediated Complement Resistance and Design Novel Therapeutics. PLoS Pathogens, 11(12), e1005290.

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