An enzyme-linked immunoassay for direct measurement of the gelatin-binding capacity of human plasma fibronectin

Damas, Pierre; Adam, Aurore; Closset, Jean; Calay, G; Lamy, Maurice; Foidart, Jean‐Michel; Mahieu, P.

doi:10.1016/0022-1759(86)90480-1

Cited by 5 publications

(7 citation statements)

References 29 publications

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“…Rabbit anti-human fibronectin antiserum (OSIE 04/ 05) was purchased from Behring (Marburg, FRG). Immunoreactive fibronectin levels were routinely measured by laser nephelometry in a Behring laser system module I-III, using the specific anti-fibronectin antiserum [13,14]. The gelatin-BC of fibronectin was measured by a solid phase ELISA.…”

Section: Measurements Offibronectin Plasma Levelsmentioning

confidence: 99%

“…The gelatin-BC of fibronectin was measured by a solid phase ELISA. The principle of this new assay has been described in detail elsewhere [13]. Briefly, wells of microwell module (NUNC, Roskilde, Denmark) were coated overnight with 200 fll of a gelatin solution of 250 ng ml " in 0·05 M carbonate buffer, pH 9·6.…”

Section: Measurements Offibronectin Plasma Levelsmentioning

confidence: 99%

“…Briefly, wells of microwell module (NUNC, Roskilde, Denmark) were coated overnight with 200 fll of a gelatin solution of 250 ng ml " in 0·05 M carbonate buffer, pH 9·6. After addition of 100 fll of 1% bovine serum albumine (BSA) for 1 h and washings with phosphate-buffered saline supplemented with 5 ml I -I of Triton x 100 (PBS-Triton) [13], the gelatin-coated wells were sequentially incubated with 100 fll of serial dilutions either of the standard plasma or of the plasma to be tested, with 100 fll of anti-human fibronectin antiserum diluted 200 times and with 100fll of horseradish peroxydase conjugated goat anti-rabbit IgG antibodies (PI 108 Behring Institute; initial dilution: 1/250). All plasmas and reagents were diluted in 0·01 M phosphate buffer, pH 7,2, containing 0·15 M NaCl (PBS) and 1% BSA.…”

Section: Measurements Offibronectin Plasma Levelsmentioning

confidence: 99%

“…We have recently set up an enzyme-linked immunoassay (ELISA) for measurement of plasma fib~onec tin concentrations [13]. Contrary to most previously described methods [14][15][16], this ELISA is based on the direct reactivity of fibronectin with gelatin.…”

Section: Introductionmentioning

confidence: 99%

“…Contrary to most previously described methods [14][15][16], this ELISA is based on the direct reactivity of fibronectin with gelatin. Furthermore it has been fully automated and appears to be as sensitive, accurate, specific and reproducible as other ELISA systems [13]. In the present work we have used a classical immunological method [14] and this ELISA to investigate the influence of Haemaccel infusion in man on both the level and the gelatin-binding capacity (gelatin-BC) of plasma fibronecti~.…”

Section: Introductionmentioning

confidence: 99%

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In‐vivo studies on Haemaccel‐fibronectin interaction in man

Damas

Adam

Buret

et al. 1987

Eur J Clin Investigation

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An enzyme-linked immunoassay has been recently set up for direct measurement of the binding capacity of plasma fibronectin to gelatin. This binding capacity could be completely inhibited in vitro by an eight-fold excess of gelatin, of Haemaccel, but not of Geloplasma. On the contrary, the levels of immunoreactive fibronectin measured by laser nephelometry did not change, in presence of 10 to 1000 micrograms ml-1 of gelatin, of Haemaccel or of Geloplasma. When infused into normal volunteers, Haemaccel provoked a strong and immediate inhibition of the plasma fibronectin binding capacity to gelatin. This inhibition was dose-dependent and maximal after infusion of 500 ml of Haemaccel. Twenty-four hours after this infusion, there was a progressive recovery of the gelatin-binding capacity, which was almost completely achieved 96 h later. The formation of complexes between Haemaccel and fibronectin was demonstrated by gel filtration chromatography and by affinity chromatography. Immunoreactive plasma fibronectin levels remained unchanged up to 24 h after infusion of 500 ml of Haemaccel. A transient decline to 50% of its initial value then occurred the second day after the infusion. Therefore, a delay existed between the formation of fibronectin-Haemaccel complexes and their elimination from the bloodstream. This delay decreased when smaller volumes of Haemaccel were infused, which strongly suggests that plasma fibronectin is cleared by means of Haemaccel and does not seem to play a role of opsonin in these conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Measurements Offibronectin Plasma Levelsmentioning

confidence: 99%

Section: Measurements Offibronectin Plasma Levelsmentioning

confidence: 99%

Section: Measurements Offibronectin Plasma Levelsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

In‐vivo studies on Haemaccel‐fibronectin interaction in man

Damas

Adam

Buret

et al. 1987

Eur J Clin Investigation

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Fibronectin: Adjunctive Therapy in Sepsis

Damas

Mahieu²,

Adam

1987

Septic Shock

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Evidence that the interaction between circulating IgA and fibronectin is a normal process enhanced in primary IgA nephropathy

et al. 1991

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A solid-phase ELISA was set up to measure the direct binding capacity (BC) of different, commercially available, purified human IgA preparations to plates coated with human fibronectin (FN). It was found that secretory, polymeric, and, to a much lesser extent, monomeric IgA exhibited elevated FN-BC as compared to their BC to plates coated with bovine serum albumin. This binding was specific since not observed with human IgG or IgM antibodies. In addition, we noted that this interaction was dose dependent, Ca2+ dependent, saturable, and not covalent, was inhibited by soluble FN, but not by a prior incubation of FN-coated plates with anti-human fibronectin antibodies, and appeared to involve on the dimeric FN other structures than its heparin-binding, collagen-binding, or C1q-binding domains. Similar experiments conducted with normal plasma indicated that plasma IgA, but not plasma IgG or IgM, was also capable of significant binding to FN-coated plates. In contrast, serum IgA did not significantly bind to those plates under otherwise identical experimental conditions. Thus, the coagulation process induces a strong decrease in the FN-BC of circulating IgA, which implies the necessity of using plasma rather than serum to study such interactions. The apparent molecular weight of plasma IgA interacting with FN-coated plates ranged between 450 and 900 kd, and its major binding characteristics were quite similar to those observed with purified polymeric IgA. The FN-BC of plasma IgA was then measured by the same ELISA in 30 patients with primary IgA nephropathy (IgAN) and in 23 healthy controls. The mean FN-BC of plasma IgA was significantly higher in patients than in normal controls. This enhancement was due mainly to the augmentation in the concentration of circulating "macromolecular" IgA and was significantly correlated with the plasma levels of IgA-FN complexes. However, the pathogenetic role of these findings was probably not determinant since similar observations were made in alcoholic liver cirrhosis without urinary abnormalities and since the FN-BC of plasma IgA or the plasma levels of IgA-FN complexes were not correlated with the various biological parameters of evolutivity of primary IgAN. In conclusion, these studies suggest that the ability of polymeric IgA to directly bind to FN is involved in the formation of circulating IgA-FN complexes and that this normal binding process, although enhanced in IgAN, is probably not responsible for kidney injury, at least in the patients studied.

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An enzyme-linked immunoassay for direct measurement of the gelatin-binding capacity of human plasma fibronectin

Cited by 5 publications

References 29 publications

In‐vivo studies on Haemaccel‐fibronectin interaction in man

In‐vivo studies on Haemaccel‐fibronectin interaction in man

Fibronectin: Adjunctive Therapy in Sepsis

Evidence that the interaction between circulating IgA and fibronectin is a normal process enhanced in primary IgA nephropathy

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