scholarly journals Development of a radioligand immunoassay for 1,25-dihydroxycholecalciferol receptors utilizing monoclonal antibody

1984 ◽  
Vol 221 (1) ◽  
pp. 129-136 ◽  
Author(s):  
S Dokoh ◽  
M R Haussler ◽  
J W Pike
Keyword(s):  
Staphylococcus Aureus ◽  
Monoclonal Antibody ◽  
D3 Receptor ◽  
Sedimentation Analysis ◽  
Binding Assays ◽  
Standard Curve ◽  
High Affinity ◽  
Cell Extracts ◽  
Mouse Myeloma

A radioligand immunoassay for 1,25-dihydroxycholecalciferol [1,25(OH)2D3] receptors was developed utilizing a specific, high-affinity (Kd = 1.8×10(-11 M) monoclonal antibody (9A7 gamma) obtained from suspension cultures of rat spleen X mouse myeloma hybrid SP2/0-9A7. A standard curve was established, based on the competition between 1,25(OH)2[3H]D3-receptor (18 fmol/tube) and increasing concentrations of radioinert 1,25(OH)2D3-receptor (0-240 fmol/tube) for the binding site on 9A7 gamma. Samples, prepared in identical buffer, contained 0-100 fmol of receptor/tube. After an equilibrium incubation of 1,25(OH)2[3H]D3-receptor with either standard or sample (16 h at 4 degrees C), antibody-bound receptor was immunoprecipitated with rabbit anti-(rat immunoglobulin) prelinked to Staphylococcus aureus and quantified. The assay is statistically sensitive to 2 fmol of receptor/tube, with intra- and inter-assay variations of 7 and 12% respectively. Occupied, unoccupied and denatured receptor were observed to compete equally in the assay. This quantitative technique has been successfully applied to the characterization of receptors after fractionation by sedimentation analysis and DNA-cellulose chromatography. Finally, the measurement of total receptor by this assay, in conjunction with 1,25(OH)2D3 binding assays, has revealed that rachitic, normal and 1,25(OH)2D3-injected chicks have respectively 13, 20, and 56% of receptor in the occupied form. From these results we consider that this radioligand immunoassay will be a useful tool in further research focusing on quantifying 1,25(OH)2D3 receptors in tissue and cell extracts.

scholarly journals Structure and characterization of a high affinity C5a monoclonal antibody that blocks binding to C5aR1 and C5aR2 receptors

mAbs ◽  
2017 ◽  
Vol 10 (1) ◽  
pp. 104-117 ◽  
Author(s):  
Caroline S. Colley ◽  
Bojana Popovic ◽  
Sudharsan Sridharan ◽  
Judit E. Debreczeni ◽  
David Hargeaves ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Affinity

Preparation Of Factor IX-Deficient Human Plasma Using RFF-IX/1 Monoclonal Antibody

1981 ◽  
Author(s):  
F Rotblat ◽  
A H Goodall ◽  
G Janossy ◽  
G Kemble ◽  
D P O’Brien ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Line ◽  
Factor Ix ◽  
Hybrid Cells ◽  
Spleen Cells ◽  
High Affinity ◽  
A Cell ◽  
A Site ◽  
Monoclonal Cell Line ◽  
Mouse Myeloma

A cell line that secretes a monoclonal antibody to factor IX has been produced by fusing spleen cells from a mouse that had been hyper immunised to purified factor IX with mouse myeloma cells (line P3-NSI/I-Ag4-1). Hybrid cells were selected and a monoclonal cell line has been established in culture. This cell line secretes an IgGl(k) antibody (RFF-IX/1) with high affinity for a site related to the coagulant function of factor IX.Monoclonal antibody was partially purified from ascitic fluid from mice implanted with the RFF-IX/1 secreting cells by precipitation at 50% saturation with ammonium sulphate. This fraction has typically 630 NIH units/ml anti IX activity and 13.5 mg/ml protein. It was coupled to cyanogen bromide activated Sepharose 2B in the ratio of 9 mg. protein/1 ml gel. A column containing 10 ml of this gel removed all the assayable factor IX from the first 280 ml of normal ci.trated plasma that was passed over it. After that volume small amounts of factor IX could be detected in the effluent. Subsequently 10-20% of the factor IX activity adsorbed could be recovered by eluting the column with 3 M potassium iodide.Immuno-affinity depleted plasma could be used as substrate in a one-stage factor IX assay under routine laboratory conditions and was undistinguishable for that purpose from severe Christmas disease plasma.

Construction and characterization of a high-affinity humanized SM5-1 monoclonal antibody

2007 ◽  
Vol 357 (4) ◽  
pp. 951-956 ◽  
Author(s):  
Bohua Li ◽  
Hao Wang ◽  
Dapeng Zhang ◽  
Weizhu Qian ◽  
Sheng Hou ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Affinity

Preparation and characterization of a high-affinity monoclonal antibody against human epididymis protein-4

2018 ◽  
Vol 141 ◽  
pp. 44-51 ◽  
Author(s):  
Yunlong Shen ◽  
Yuxi Wang ◽  
Xiaohua Jiang ◽  
Liang Lu ◽  
Chengdi Wang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Human Epididymis Protein 4 ◽  
High Affinity ◽  
Human Epididymis

Production and characterization of murine monoclonal antibodies against staphylococcal enterotoxins A and E

1991 ◽  
Vol 37 (8) ◽  
pp. 581-585 ◽  
Author(s):  
Kunihiro Shinagawa ◽  
Emiko Nishimura ◽  
Makoto Mitsumori ◽  
Naonori Matsusaka ◽  
Shunji Sugii
Keyword(s):  
Staphylococcus Aureus ◽  
Monoclonal Antibodies ◽  
The Other ◽  
Staphylococcal Enterotoxin A ◽  
Binding Assays ◽  
Spleen Cells ◽  
Myeloma Cells ◽  
Murine Monoclonal Antibodies ◽  
Competitive Binding Assays

Six murine monoclonal antibodies (MAbs) against staphylococcal enterotoxin A (SEA) and enterotoxin E (SEE) were prepared by fusion of myeloma cells with mouse spleen cells immunized with SEA and SEE. Of five MAbs to SEA tested, two MAbs were reactive with only SEA, whereas three were specific for both SEA and SEE. On the other hand, one MAb to SEE was found to be specific for only SEE. To study specificities of the combining sites of these MAbs, competitive binding assays with either SEA or SEE and horseradish peroxidase conjugated MAbs were performed using unconjugated MAbs as inhibitors. The results obtained in the assays suggest that different epitopes may be located on SEA and that some of them may be cross-reacting epitopes between SEA and SEE. Key words: enterotoxins, monoclonal antibodies, Staphylococcus aureus.

scholarly journals Characterization of a Humanized Monoclonal Antibody Recognizing Clumping Factor A Expressed by Staphylococcus aureus

2005 ◽  
Vol 73 (8) ◽  
pp. 5229-5232 ◽  
Author(s):  
Paul J. Domanski ◽  
Pratiksha R. Patel ◽  
Arnold S. Bayer ◽  
Li Zhang ◽  
Andrea E. Hall ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Monoclonal Antibody ◽  
Infective Endocarditis ◽  
Binding Kinetics ◽  
Vancomycin Therapy ◽  
Latex Beads ◽  
Humanized Monoclonal Antibody ◽  
Kinetics Of ◽  
Therapeutic Model

ABSTRACT We report the humanization and characterization of monoclonal antibody (MAb) T1-2 or tefibazumab, a monoclonal antibody that recognizes clumping factor A expressed on the surface of Staphylococcus aureus. We demonstrate that the binding kinetics of MAb T1-2 is indistinguishable compared to that of its murine parent. Furthermore, MAb T1-2 is shown to enhance the opsonophagocytic uptake of ClfA-coated latex beads, protect against an intravenous challenge in a prophylactic model of rabbit infective endocarditis, and enhance the efficacy of vancomycin therapy in a therapeutic model of established infective endocarditis.

Preparation and characterization of a high-affinity monoclonal antibody against nerve growth factor

2021 ◽  
pp. 105966
Author(s):  
Shuang Liu ◽  
Yunlong Shen ◽  
Pengyu Chen ◽  
Cuiyu Guo ◽  
Guangbing Zhang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Nerve Growth ◽  
High Affinity
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