Interaction of monoclonal antibodies with growth hormone-binding protein and its complex with growth hormone

1993 ◽  
Vol 139 (3) ◽  
pp. 495-501 ◽  
Author(s):  
H. Sadeghi ◽  
A. L. Lumanglas ◽  
W. R. Baumbach ◽  
B. S. Wang
Keyword(s):  
Growth Hormone ◽  
Monoclonal Antibodies ◽  
Binding Assay ◽  
Interaction Analysis ◽  
Binding Protein ◽  
Antigenic Determinants ◽  
Binding Assays ◽  
Competition Binding Assay ◽  
Competition Binding ◽  
Sequential Binding

ABSTRACT The properties of four independent lines of monoclonal antibodies (MAbs) specific to rat GH-binding protein (GHBP) were examined. Three MAbs, designated GHR-12, GHR-13 and GHR-16, were raised against the entire GHBP molecule. The fourth MAb, designated as GHBP4·3, was raised against the 17 amino acid residues at the C-terminal end of rat GHBP. The interaction of these antibodies with GHBP and their effect on GH binding to GHBP were analysed by conventional competition binding assays and surface plasmon resonance, i.e. with a Biospecific Interaction Analysis (BIAcore) instrument. The binding affinity of these MAbs to GHBP ranged from 29 nmol/l to 30·9 pmol/l. The pair-wise antibody binding to GHBP on BIAcore suggested that GHR-13 and GHR-16 recognized different antigenic determinants while part of the GHR-12 epitope might be shared with the other antibodies. The antibodies inhibited the interaction of GH with GHBP in the competition binding assay. However, in sequential binding on the BIAcore instrument, they were able to bind GHBP after its interaction with GH, indicating that the inhibition observed in the competition binding assay resulted from steric hindrance rather than direct interference with the GH-binding site of GHBP. The present findings, therefore, suggest that these antibodies are useful for investigating GHBP and its interaction with GH. Journal of Endocrinology (1993) 139, 495–501

scholarly journals Comparisons of rotavirus VP7-typing monoclonal antibodies by competition binding assay.

1992 ◽  
Vol 30 (3) ◽  
pp. 704-711 ◽  
Author(s):  
P Raj ◽  
D O Matson ◽  
B S Coulson ◽  
R F Bishop ◽  
K Taniguchi ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Binding Assay ◽  
Competition Binding Assay ◽  
Competition Binding

scholarly journals Circumsporozoite proteins of malaria parasites contain a single immunodominant region with two or more identical epitopes.

1983 ◽  
Vol 157 (6) ◽  
pp. 1947-1957 ◽  
Author(s):  
F Zavala ◽  
A H Cochrane ◽  
E H Nardin ◽  
R S Nussenzweig ◽  
V Nussenzweig
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Specific Binding ◽  
Polyclonal Antibodies ◽  
Binding Assays ◽  
Malaria Parasites ◽  
Immunodominant Region ◽  
Additional Finding ◽  
Single Region ◽  
Competition Binding

We have used panels of monoclonal antibodies to circumsporozoite (CS) proteins of Plasmodium falciparium, P. vivax, and P. knowlesi to determine the number of topographically independent epitopes of these antigens. The results of competition binding assays indicated that single regions of the CS molecules were recognized by the homologous monoclonal antibodies. Competition binding assays were also used to study the specificity of antibodies contained in the sera of humans and monkeys that had developed sterile immunity after immunization with irradiated, intact sporozoites. We found that single monoclonal antibodies inhibited 70-95% of the specific binding of the polyclonal antibodies to crude extracts of sporozoites. It appears, therefore, that CS proteins are among the most immunogenic constituents of sporozoites, and that a single region of these molecules contains most of the immunogenic activity. An additional finding was that the immunodominant region of CS molecules is multivalent with regard to the expression of a single epitope. This was demonstrated by the ability of monomers of CS proteins to bind simultaneously two or more molecules of the same monoclonal antibody.

scholarly journals A competition binding assay for determination of the inositol (1,4,5)-trisphosphate content of human leucocytes

1990 ◽  
Vol 170 (2) ◽  
pp. 755-762 ◽  
Author(s):  
Peter H. Nibbering ◽  
Timo P.L. Zomerdijk ◽  
Peter J.M. van Haastert ◽  
Ralph van Furth
Keyword(s):  
Binding Assay ◽  
Competition Binding Assay ◽  
Inositol 1,4,5 Trisphosphate ◽  
Competition Binding

scholarly journals Effect of immunization of hamsters against recombinant P26h on fertility rates

Reproduction ◽  
2002 ◽  
pp. 307-313 ◽  
Author(s):  
C Gaudreault ◽  
L Montfort ◽  
R Sullivan
Keyword(s):  
Zona Pellucida ◽  
Binding Protein ◽  
Maltose Binding Protein ◽  
Fertilization Rate ◽  
Active Immunization ◽  
Antigenic Determinants ◽  
Binding Assays ◽  
Contraceptive Vaccine

Despite the various contraceptive methods available, an effective and inexpensive method remains to be established. Immunocontraception may help to achieve this goal. P26h has been proposed as a candidate for the development of a male contraceptive vaccine. P26h, a hamster sperm protein, interacts with the zona pellucida. Furthermore, in vivo fertilization can be blocked completely by active immunization of male hamsters against P26h. Maltose binding protein (MBP)-P26 shares antigenic determinants with the native P26h present on cauda epididymal spermatozoa. The aim of the present study was to reproduce the immunocontraceptive properties of native P26h by immunizing male hamsters against a recombinant P26h fused with a maltose binding protein (MBP). Active immunization of male hamsters with the MBP-P26h showed that specific anti-P26h circulating IgGs could be generated. Mating of immunized male hamsters with superovulated females resulted in a significant decrease, 20-25%, in the fertilization rate. This result is in agreement with results from in vitro sperm-zona pellucida binding assays. Indeed, the anti-recombinant P26h IgGs showed lower inhibitory properties when compared with anti-native P26h IgG. Despite the high anti-P26h IgG titres generated in hamsters, histological studies showed that active immunization has no pathological sequelae to the reproductive tissues. The potential of P26h as a candidate for a contraceptive vaccine is discussed.

Detection and quantitation of amanitin using an RNA-polymerase competition binding assay

Toxicon ◽  
1980 ◽  
Vol 18 (5-6) ◽  
pp. 702-704
Author(s):  
Arnold Brown ◽  
George M. Garrity
Keyword(s):  
Rna Polymerase ◽  
Binding Assay ◽  
Competition Binding Assay ◽  
Competition Binding

Fluorescence polarization-based competition binding assay for c-Jun N-terminal kinases 1 and 2

2017 ◽  
Vol 532 ◽  
pp. 26-28 ◽  
Author(s):  
Francesco Ansideri ◽  
Marcel Dammann ◽  
Frank M. Boeckler ◽  
Pierre Koch
Keyword(s):  
Fluorescence Polarization ◽  
Binding Assay ◽  
Competition Binding Assay ◽  
Competition Binding

scholarly journals Discovery of BAZ2A Bromodomain Ligands

2017 ◽  
Author(s):  
Dimitrios Spiliotopoulos ◽  
Eike-Christian Wamhoff ◽  
Graziano Lolli ◽  
Christoph Rademacher ◽  
Amedeo Caflisch
Keyword(s):  
Small Molecules ◽  
Binding Assay ◽  
Side Chain ◽  
Binding Modes ◽  
Zinc Finger Domain ◽  
Ligand Efficiency ◽  
Competition Binding Assay ◽  
Energy Evaluation ◽  
Natural Ligand ◽  
Competition Binding

The bromodomain adjacent to zinc finger domain protein 2A (BAZ2A) is implicated in aggressive prostate cancer. The BAZ2A bromodomain is a challenging target because of the shallow pocket of its natural ligand, the acetylated side chain of lysine. Here, we report the successful screening of a library of nearly 1500 small molecules by high-throughput docking and force field-based binding-energy evaluation. For seven of the 20 molecules selected in silico, evidence of binding to the BAZ2A bromodomain is provided by ligand-observed NMR spectroscopy. Two of these compounds show a favorable ligand efficiency of 0.42 kcal/mol per non-hydrogen atom in a competition-binding assay. The crystal structures of the BAZ2A bromodomain in complex with four fragment hits validate the predicted binding modes. The binding modes of compounds 1 and 3 are compatible with ligand growing for optimization of affinity for BAZ2A and selectivity against the close homologue BAZ2B.

scholarly journals Development of a Competition-Binding Assay to Determine Binding Affinity of Molecules to Neuromelanin via Fluorescence Spectroscopy

Biomolecules ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 175 ◽  
Author(s):  
Jackson Fink ◽  
Heather Pathak ◽  
John Smith ◽  
Cindy Achat-Mendes ◽  
Robert L. Haining
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Binding Affinity ◽  
Dopaminergic Neurons ◽  
Binding Assay ◽  
Pathological State ◽  
Polymeric Form ◽  
Competition Binding Assay ◽  
Competition Binding ◽  
6 Hydroxydopamine

Neuromelanin, the polymeric form of dopamine which accumulates in aging neuronal tissue, is increasingly recognized as a functional and critical component of a healthy and active adult human brain. Notorious in plant and insect literature for their ability to bind and retain amines for long periods of time, catecholamine polymers known colloquially as ‘melanins’ are nevertheless curiously absent from most textbooks regarding biochemistry, neuroscience, and evolution. Recent research has brought attention to the brain pigment due to its possible role in neurodegeneration. This linkage is best illustrated by Parkinson’s disease, which is characterized by the loss of pigmented dopaminergic neurons and the ‘white brain’ pathological state. As such, the ability to determine the binding affinity of neurotoxic agents, as well as any potential specific endogenous ligands to neuromelanin are of interest and potential value. Neuromelanin has been shown to have saturable binding interactions with nicotine as monitored by a fluorimeter. This interaction provides a signal to allow for a competition-binding assay with target molecules which do not themselves produce signal. The current report establishes the viability of this competition assay toward three compounds with central relevance to Parkinson’s disease. The Kd of binding toward neuromelanin by methyl-phenyl-pyridinium ion (MPP+), dopamine, and 6-hydroxydopamine were found to be 1 mM, 0.05 mM, and 0.1 mM, respectively in the current study. In addition, we demonstrate that 6-hydroxydopamine polymerizes to form neuromelanin granules in cultured dopaminergic neurons that treated with 2,4,5-trihydroxy-l-phenylalanine. Immunohistochemical analysis using fluor-tagged anti-dopamine antibodies suggests that the incorporation of 6-hydroxydopamine (following internalization and decarboxylation analogous to levodopa and dopamine) alters the localized distribution of bound dopamine in these cells.

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