Specific Precipitating Antibodies To VIII: CAg In Two Patients With Haemophilia

1981 ◽  
Author(s):  
J M Lavergne ◽  
D Meyer ◽  
J M Girma ◽  
M J Larrieu
Keyword(s):  
Lattice Theory ◽  
Solid Phase ◽  
Ionic Composition ◽  
Double Diffusion ◽  
Sodium Thiocyanate ◽  
Haemophilia A ◽  
Precipitin Line ◽  
Fab Fragments ◽  
Normal Plasma ◽  
Highly Sensitive

IgG and Fab fragments were prepared from the plasma of two haemophilia A patients whose anti- V111 : C titres were 2,000 and 400 Bethesda U./ml. 125-I-IgG and Fab specific for VIII :CAg were purified by a solid phase procedure and the reactivity of these two antibodies with VIII: CAg was studied by immunoprecipitation in agarose. With double diffusion and autoradiographic methods, both the IgG and Fab antibodies showed a precipitin line against normal plasma, serum, haemophilia A+ plasma, cryopre ci pi t ate , purified F.VIII/vWF or VIII: C free of vWF. No precipitin line was observed in haemophilia A- or severe von Willebrand’s disease. With electroimmunoassay (radi o-Laurell) using both IgG and Fab antibodies, results of VIII: C A g were in agreement in all samples with those obtained by immunoradiometric assay using the same antibodies. The specificity of the immunoprecipitation observed in agarose with IgG or Fab fragments was assessed by modifying the pH (7.5 to 9.5) of the buffer, the ionic composition (0.15, 0.5 and 1M sodium chloride, 0.15 and 0.5M potassium iodide, 0.15 and 0.5M sodium thiocyanate) of the washing fluid,or by carbamy1ation of the anti-VIII: CAg IgG. In all cases, specific precipitation was observed. In addition, 125-I-IgG or Fab isolated by the same technique from normal plasma gave no precipitinline with VIII: CAg. This study demonstrates that, contrary to previous evidence, 1) human anti-VIII: CAg antibodies are both precipitating as well as neutralizing when studied by highly sensitive techniques; and 2) monovalent Fab can also precipitate VIII: C Ag. The results of this study raise some questions about the lattice theory of immunoprecipitation.

Highly Sensitive Visual Fluorometry of Aluminium at ppb Level with Ring-like Solid Phase of Poly(vinyl alcohol)

1999 ◽  
Vol 28 (3) ◽  
pp. 217-218 ◽  
Author(s):  
Akihiko Ishida ◽  
Emiko Kaneko ◽  
Takao Yotsuyanagi
Keyword(s):  
Vinyl Alcohol ◽  
Solid Phase ◽  
Poly Vinyl Alcohol ◽  
Highly Sensitive

scholarly journals A monoclonal anti-human plasma prekallikrein antibody that inhibits activation of prekallikrein by factor XIIa on a surface

Blood ◽  
1987 ◽  
Vol 70 (4) ◽  
pp. 1053-1062
Author(s):  
D Veloso ◽  
LD Silver ◽  
S Hahn ◽  
RW Colman
Keyword(s):  
Heavy Chain ◽  
Sodium Dodecyl ◽  
Factor Xii ◽  
Contact Activation ◽  
Fab Fragments ◽  
Microtiter Plates ◽  
Normal Plasma ◽  
Reducing Conditions ◽  
Factor Xiia ◽  
Kallikrein Activity

Of five IgGI/k murine monoclonal anti-human prekallikrein antibodies produced (MAbs), MAb 13G11 was selected for studying interaction of prekallikrein with factor XII and high-mol-wt kininogen (HMWK) during activation on a surface. Immunoblots from sodium dodecyl sulfate (SDS) gels showed that this MAb recognizes two variants (88 kd and 85 kd) of prekallikrein and kallikrein both in purified proteins and normal plasma. Under reducing conditions, kallikrein exhibits the epitope on the heavy chain but not on the light chains. Preincubation of MAb 13G11 with prekallikrein (added to prekallikrein-deficient plasma) or with normal plasma inhibited surface activation of prekallikrein 60% to 80%, as judged by amidolytic and coagulant assays. In normal plasma, inhibition by the Fab fragments was 87% of that with the entire MAb. Inhibition was not by competition between the MAb and HMWK, since neither binding of 13G11 to prekallikrein (coated on microtiter plates) was inhibited by an excess of HMWK, nor was hydrolysis of HMWK by kallikrein inhibited by 13G11. Using purified proteins in a system mimicking contact activation, inhibition by 13G11 of prekallikrein activation by factor XIIa, HMWK, and kaolin present was approximately 80%. Decreased inhibition (55% to 25%) occurred without HMWK or when kallikrein was used instead of prekallikrein. Kallikrein activity was not inhibited by 13G11 Fab fragments. These results indicate that the effect of 13G11 in plasma was neither dissociation of prekallikrein- HMWK complex nor a direct effect on kallikrein activity. Similar to the results in plasma, activation of prekallikrein, HMWK present, by factor XIIa bound to kaolin, was inhibited approximately 70% by 13G11. The results suggest a previously unrecognized site on the prekallikrein (heavy chain) required for its interaction with factor XIIa, either shared with the 13G11 epitope or located in very close proximity. The inhibition of kallikrein by intact 13G11 indicates that its binding site on the heavy chain is sterically related to the active site (light chain).

scholarly journals Highly sensitive non-enzymatic hydrogen peroxide monitoring platform based on nanoporous gold via a modified solid-phase reaction method

RSC Advances ◽  
2021 ◽  
Vol 11 (58) ◽  
pp. 36753-36759
Author(s):  
Zhipeng Yang ◽  
Jun Li ◽  
Panmei Liu ◽  
An Zhang ◽  
Jing Wang ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Solid Phase ◽  
Nanoporous Gold ◽  
Solid Phase Reaction ◽  
Selective Detection ◽  
Phase Reaction ◽  
Reaction Method ◽  
Grain Sizes ◽  
Highly Sensitive ◽  
Monitoring Platform

Ge/Au/Ge triple-layered precursor was proposed to prepare nanoporous gold (NPG) with much smaller grain sizes and nanopores as an electrochemical sensor for highly sensitive and selective detection of hydrogen peroxide.

scholarly journals Nano-ion-exchangers - a new class of reactive materials

2018 ◽  
Vol 18 (6) ◽  
pp. 794-809
Author(s):  
Anatoly M. Dolgonosov ◽  
Ruslan Kh. Khamizov ◽  
Nadezhda K. Kolotilina
Keyword(s):  
Solid Phase ◽  
Ionic Composition ◽  
Photo Luminescence ◽  
Colloidal Systems ◽  
Ion Exchangers ◽  
New Class ◽  
Counter Ions ◽  
Dynamic Experiments ◽  
Back Ground ◽  
Charged Ions

Nano-sized particles of functional polymers i.e. nano-ion-exchangers (NIEX), are unusual objects which simultaneously behave as the hyper-charged ions and the solid ion exchangers. Due to similar charges, they form very stable colloidal systems. This paper is devoted to theoretical and practical study of the proper- ties of nano-exchangers, methods for their preparation, the technique of experiments being practically un- known, and the opportunities for their application. The results of dynamic experiments are given for sorption of nano-exchangers and the ions of back- ground solutions on the beds of macro-particles of usual cationic and anionic resins. It is shown how to ob- tain the NIEX hydrosols with the desired ionic composition, and the concept of the standard state hydrosol is defined. The possibility for solid-phase exchange of counter ions between contacting particles of the same polarity is demonstrated. The possibilities and advantages of using nano-ion-exchangers in chemical analysis are demonstrat- ed by different examples: preparation of separating phases for ion chromatography, application as modifier in capillary electrophoresis and using in photo-luminescence. The prospects of nano-ion-exchangers for drug delivery are also shown.

scholarly journals A monoclonal anti-human plasma prekallikrein antibody that inhibits activation of prekallikrein by factor XIIa on a surface

Blood ◽  
1987 ◽  
Vol 70 (4) ◽  
pp. 1053-1062 ◽  
Author(s):  
D Veloso ◽  
LD Silver ◽  
S Hahn ◽  
RW Colman
Keyword(s):  
Heavy Chain ◽  
Sodium Dodecyl ◽  
Factor Xii ◽  
Contact Activation ◽  
Fab Fragments ◽  
Microtiter Plates ◽  
Normal Plasma ◽  
Reducing Conditions ◽  
Factor Xiia ◽  
Kallikrein Activity

Abstract Of five IgGI/k murine monoclonal anti-human prekallikrein antibodies produced (MAbs), MAb 13G11 was selected for studying interaction of prekallikrein with factor XII and high-mol-wt kininogen (HMWK) during activation on a surface. Immunoblots from sodium dodecyl sulfate (SDS) gels showed that this MAb recognizes two variants (88 kd and 85 kd) of prekallikrein and kallikrein both in purified proteins and normal plasma. Under reducing conditions, kallikrein exhibits the epitope on the heavy chain but not on the light chains. Preincubation of MAb 13G11 with prekallikrein (added to prekallikrein-deficient plasma) or with normal plasma inhibited surface activation of prekallikrein 60% to 80%, as judged by amidolytic and coagulant assays. In normal plasma, inhibition by the Fab fragments was 87% of that with the entire MAb. Inhibition was not by competition between the MAb and HMWK, since neither binding of 13G11 to prekallikrein (coated on microtiter plates) was inhibited by an excess of HMWK, nor was hydrolysis of HMWK by kallikrein inhibited by 13G11. Using purified proteins in a system mimicking contact activation, inhibition by 13G11 of prekallikrein activation by factor XIIa, HMWK, and kaolin present was approximately 80%. Decreased inhibition (55% to 25%) occurred without HMWK or when kallikrein was used instead of prekallikrein. Kallikrein activity was not inhibited by 13G11 Fab fragments. These results indicate that the effect of 13G11 in plasma was neither dissociation of prekallikrein- HMWK complex nor a direct effect on kallikrein activity. Similar to the results in plasma, activation of prekallikrein, HMWK present, by factor XIIa bound to kaolin, was inhibited approximately 70% by 13G11. The results suggest a previously unrecognized site on the prekallikrein (heavy chain) required for its interaction with factor XIIa, either shared with the 13G11 epitope or located in very close proximity. The inhibition of kallikrein by intact 13G11 indicates that its binding site on the heavy chain is sterically related to the active site (light chain).

Highly sensitive assays of autoantibodies to thyroglobulin and to thyroid peroxidase.

1989 ◽  
Vol 35 (9) ◽  
pp. 1949-1954 ◽  
Author(s):  
K Beever ◽  
J Bradbury ◽  
D Phillips ◽  
S M McLachlan ◽  
C Pegg ◽  
...  
Keyword(s):  
Solid Phase ◽  
Protein A ◽  
Cross Reactivity ◽  
Thyroid Peroxidase ◽  
Scatchard Analysis ◽  
Thyroid Autoantibodies ◽  
Serum Samples ◽  
Highly Sensitive ◽  
Phase Protein ◽  
High Concentrations

Abstract These highly sensitive assays are based on the interaction between thyroid autoantibodies and 125I-labeled autoantigens. Serum samples are incubated with labeled thyroid peroxidase (TPO) or thyroglobulin (Tg) to allow the formation of antibody-labeled antigen complexes. The complexes are then precipitated by addition of solid-phase Protein A. In the presence of high concentrations of TPO antibody or Tg antibody, more than 50% of the respective labeled antigen was precipitated, whereas only 1-2% was precipitated in the absence of autoantibody. Interassay CVs were 3.2% and 5.7%, respectively, for the anti-TPO and anti-Tg assays. There was no cross-reactivity between Tg antibody and TPO antibody. Results correlated highly significantly with results from other assay systems based on antigen-coated cells or plastic supports, but the assays described here were considerably more sensitive. Scatchard analysis of the assay data provided information on the affinity and serum concentration of TPO autoantibodies (ka approximately 10(9) L/mol and concentrations up to 1 g/L) and Tg autoantibodies (ka approximately 4 x 10(10) L/mol and concentrations up to 1 g/L). Overall, these assays provide a sensitive, precise, and convenient system for measuring and investigating the properties of thyroid autoantibodies.

Start of UK Confidential Haemophilia A Database: Analysis of 142 Patients by Solid Phase Fluorescent Chemical Cleavage of Mismatch

1999 ◽  
Vol 81 (06) ◽  
pp. 900-905 ◽  
Author(s):  
Naushin Waseem ◽  
Richard Bagnall ◽  
Peter Green ◽  
Francesco Giannelli ◽  
Keyword(s):  
Factor Viii ◽  
Solid Phase ◽  
Haemophilia A ◽  
National Strategy ◽  
Coding Region ◽  
Database Analysis ◽  
Chemical Cleavage ◽  
Signal Region ◽  
The Uk ◽  
Von Willebrand

SummaryA national strategy for optimising genetic services in haemophilia A has been initiated in the UK. Solid phase fluorescent chemical cleavage of mismatch is used to screen the entire coding region of factor VIII in six segments: four amplified from the trace of mRNA in blood lymphocytes and two from genomic DNA for the 3.4 kb exon 14 and flanking intron sequences. These segments are analysed in two threefold multiplexes so that the genes of 18 patients can be screened in a single ABI 377 gel. The promoter and polyadenylation signal region are amplified and sequenced directly. We have analysed 142 unrelated patients and identified 141 factor VIII mutations and one Normandy type von Willebrand homozygote. The former mutations include 89 missense, 10 nonsense, 5 frameshift, one 24 bp deletion and one splice signal defect. These comprise 71 different changes, of which 39 have not been previously observed.

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