A monoclonal antibody to C1q which appears to interact with C½C½s<sub>2</sub>‐binding site

Hsiung, Li-min; Dodds, Alister W.; Mason, Donald W.

doi:10.1016/0014-5793(88)80789-0

Cited by 4 publications

(1 citation statement)

References 24 publications

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“…So far, MAbs to human complement components have been raised by immunizing mice with purified complement factors (24,25,26). Monoclonal antibodies to mouse complement have been obtained in the case of C5 by immunizing C5 deficient mice with serum from normal mice (27).…”

Section: Discussionmentioning

confidence: 99%

Monoclonal Antibodies to Complement Components Without the Need of Their Prior Purification. II. Antibodies to Mouse C3 And C4

Kremmer¹,

Thierfelder²,

Felber³

et al. 1990

Hybridoma

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Rat monoclonal antibodies (MAbs) to mouse complement components C3 and C4 were produced by immunizing rats with cell-bound C3 and C4. This principle involves: a) using mouse thymocytes coated with syngeneic rat antibody isotypes that show high affinity to C1q, b) the intercalation of C1q from serum and c) the subsequent activation of the classical complement pathway leading to deposition of cell-bound complement components. Screening for anti-complement antibodies was performed on antibody coating microtiter plates with mouse serum as source of complement. The reactivity of the MAbs was determined by variations of the ELISA screening system using EDTA-serum to inhibit complement activation by C1 dissociation, serum rendered deficient of functionally active C3 by treatment with cobra venom factor (CVF) or serum of genetically C5-deficient mice. The specificity of the MAbs was confirmed by affinity chromatography followed by SDS-PAGE and immunoblotting. We were able to establish a panel of anti-C3 and anti-C4 MAbs of various isotypes.

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Section: Discussionmentioning

confidence: 99%

Monoclonal Antibodies to Complement Components Without the Need of Their Prior Purification. II. Antibodies to Mouse C3 And C4

Kremmer¹,

Thierfelder²,

Felber³

et al. 1990

Hybridoma

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Chemical characterization and location of ionic interactions involved in the assembly of the C1 complex of human complement

1993

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The C1 complex of human complement comprises two loosely interacting subunits, C1q and the Ca(2+)-dependent C1s-C1r-C1r-C1s tetramer. With a view to gain information on the nature of the ionic interactions involved in C1 assembly, we have studied the effects of the chemical modifications of charged residues of C1q or the tetramer on their ability to reconstitute the C1 complex. Treatment of C1q with pyridoxal-5'-phosphate, acetic anhydride, and citraconic anhydride, as well as with cyclohexanedione and diethylpyrocarbonate, inhibited its ability to associate with C1s-C1r-C1r-C1s. Treatment of the collagen-like fragments of C1q with the same reagents yielded the same effects. Treatment of C1s-C1r-C1r-C1s with 1-ethyl-3-[-3-(dimethylamino) propyl] carbodiimide also prevented C1 assembly, through modification of acidic amino acids which were shown to be located in C1r. Further studies on the location of the interaction sites within C1q, using ligand-blotting and competition experiments with synthetic peptides, were unsuccessful, suggesting that these sites are contributed to by two or three of the C1q chains. It is concluded that C1 assembly involves interactions between acidic amino acids of C1r and lysine (hydroxylysine) and arginine residues located within the collagen-like region of C1q. Sequence comparison with mannan binding protein, another collagen-like molecule which binds the C1s-C1r-C1r-C1s tetramer, suggests Arg A38, and HyL B32, B65, and C29 of C1q as possible interaction sites.

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Analysis of Human C1q by Combined Bottom-up and Top-down Mass Spectrometry

Pflieger

Przybylski

Gonnet

et al. 2010

Molecular & Cellular Proteomics

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C1q is a subunit of the C1 complex, a key player in innate immunity that triggers activation of the classical complement pathway. Featuring a unique structural organization and comprising a collagen-like domain with a high level of post-translational modifications, C1q represents a challenging protein assembly for structural biology. We report for the first time a comprehensive proteomics study of C1q combining bottom-up and top-down analyses.

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A monoclonal antibody to C1q which appears to interact with C½C½s₂‐binding site

Cited by 4 publications

References 24 publications

Monoclonal Antibodies to Complement Components Without the Need of Their Prior Purification. II. Antibodies to Mouse C3 And C4

Monoclonal Antibodies to Complement Components Without the Need of Their Prior Purification. II. Antibodies to Mouse C3 And C4

Chemical characterization and location of ionic interactions involved in the assembly of the C1 complex of human complement

Analysis of Human C1q by Combined Bottom-up and Top-down Mass Spectrometry

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A monoclonal antibody to C1q which appears to interact with C½C½s2‐binding site

Cited by 4 publications

References 24 publications

Monoclonal Antibodies to Complement Components Without the Need of Their Prior Purification. II. Antibodies to Mouse C3 And C4

Monoclonal Antibodies to Complement Components Without the Need of Their Prior Purification. II. Antibodies to Mouse C3 And C4

Chemical characterization and location of ionic interactions involved in the assembly of the C1 complex of human complement

Analysis of Human C1q by Combined Bottom-up and Top-down Mass Spectrometry

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A monoclonal antibody to C1q which appears to interact with C½C½s₂‐binding site