Stepwise “bridge-to-bridge” reduction of monoclonal antibodies and light chain detection: Case studies of tenatumomab and trastuzumab

2018 ◽  
Vol 1 (4) ◽  
pp. 261-269 ◽  
Author(s):  
Giulia Mazzoccanti ◽  
Giuseppe Pierri ◽  
Alessia Ciogli ◽  
Omar H. Ismail ◽  
Fabrizio Giorgi ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Case Studies ◽  
Light Chain

Back Cover: Stepwise “bridge-to-bridge” reduction of monoclonal antibodies and light chain detection: Case studies of tenatumomab and trastuzumab

2018 ◽  
Vol 1 (4) ◽  
pp. NA-NA
Author(s):  
Giulia Mazzoccanti ◽  
Giuseppe Pierri ◽  
Alessia Ciogli ◽  
Omar H. Ismail ◽  
Fabrizio Giorgi ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Case Studies ◽  
Light Chain ◽  
Back Cover

Preferential light-chain labeling of native monoclonal antibodies improves the properties of fluorophore conjugates

2021 ◽  
pp. 153211
Author(s):  
Michael P. Luciano ◽  
Ivan Dingle ◽  
Saghar Nourian ◽  
Martin J. Schnermann
Keyword(s):  
Monoclonal Antibodies ◽  
Light Chain

Molecular Characterization of Variable Heavy and Light Chain Regions of Five HIV Type 1-Specific Human Monoclonal Antibodies*

1994 ◽  
Vol 10 (12) ◽  
pp. 1639-1649 ◽  
Author(s):  
ERIC M.M. van der DONK ◽  
MARTIN SCHUTTEN ◽  
ALBERT D.M.E. OSTERHAUS ◽  
ROGER W.J. van der HEIJDEN
Keyword(s):  
Monoclonal Antibodies ◽  
Molecular Characterization ◽  
Light Chain ◽  
Human Monoclonal Antibodies ◽  
Variable Heavy ◽  
Hiv Type 1

MONOCLONAL ANTIBODIES THAT RECOGNIZE Ca2+-INDUCED CONFORMER OF PROTEIN C, INDEPENDENT OF GLA RESIDUES

1987 ◽  
Author(s):  
T Sugo ◽  
S Tanabe ◽  
K Shinoda ◽  
M Matsuda
Keyword(s):  
Monoclonal Antibodies ◽  
Light Chain ◽  
Metal Binding ◽  
Binding Sites ◽  
Plasma Proteins ◽  
Protein C ◽  
The Other ◽  
Metal Binding Sites ◽  
Direct Elisa ◽  
Gla Domain

Monoclonal antibodies (MCA’s) were prepared against human protein C (PC) according to Köhler & Milstein, and those that recognize the Ca2+-dependent PC conformers were screened by direct ELISA in the presence of 2 mM either CaCl2 or EDTA. Out of nine MCAߣs thus screened, five MCA's designated as HPC-1˜5, respectively, were found to react with PC in the presence of Ca2+ but not EDTA. By SDS-PAGE coupled with Western Blotting performed in the presence of 2 mM CaCl2, we found that two MCA’s HPC-1 and 2, recognized the light chain, and two others, HPC-3 and 4, recognized the heavy chain of PC. But another MCA, HPC-5 was found to react with only non-reduced antigens. Further study showed that HPC-1 and 5 failed to react with the Gla-domainless PC, i.e. PC from which the N-terminal Gla-domain of the light chain had been cleaved off by α-chymotrypsin. However, all the other three MCA's retained the reactivity with the antigen in the presence of Ca2+ even after the Gla-domain had been removed. The binding of these MCA’s to PC in the presence of Ca2+ was found to be saturable with respect to the Ca2+ concentration and the half maximal binding for each MCA was calculated to be about 0.5+mM. Moreover, many other divalent cations such as Mg2+, Mn2+ , Ba2+, Zn2+, Co2+, Sr2+, were found to substitute for Ca2+ in inducing the metal ion-dependent but Gla-domain-independent conformer of PC.Cross-reactivity to other vitamin K-aependent plasma proteins was examined by direct ELISA; HPC-2 and 3 reacted solely to PC, but HPC-1 and 4 also reacted with prothrombin and HPC-5 with both prothrombin and factor X.These findings indicated that there are two or more metal binding sites besides the Gla-domain, possibly one in the light chain and the other(s) in the heavy chain. The presence of these metal binding sites may contribute to the unique conformer of vitamin K-dependent plasma proteins including protein C.

F VIII Subunits: Purification and Antigenic Properties

1987 ◽  
Vol 58 (04) ◽  
pp. 1043-1048 ◽  
Author(s):  
Ole Nordfang ◽  
Mirella Ezban ◽  
Jan J Hansen
Keyword(s):  
Monoclonal Antibodies ◽  
Heavy Chain ◽  
Light Chain ◽  
Hemophilia A ◽  
Exchange Chromatography ◽  
Inhibition Assay ◽  
Cation Exchange Chromatography ◽  
Inhibitor Activity ◽  
Fviii Inhibitor ◽  
Antigenic Properties

SummaryFactor VIII-Light Chain (FVIII-LC) and FVIII-Heavy Chain (FVIII-HC) were purified from human plasma by the use of immunosorbents containing monoclonal antibodies or human inhibitor antibodies. FVIII-LC was subsequently isolated in essentially pure state by cation exchange chromatography. The preparations obtained contained 50 ng of protein for each unit of FVIII-LC antigen (FVIII-LC: Ag).Affinity purified FVIII-LC and FVIII-HC preparations containing less than 0.3% of the opposite subunit were added in FVIILC inhibition assay of hemophilia A inhibitor antibodies. FVIII-LC was able to fully block the inhibitor activity in 6 out of 7 hemophilia A plasmas and partially block the inhibitor activity of one plasma. FVIII-HC only blocked FVIILC inhibiting antibodies form the plasma that was not fully blocked by FVIII-LC. It is suggested that FVIII-LC can be used for immunotherapy of the patients whose FVIILC inhibiting antibodies are directed towards FVIII-LC.When FVIII-LC was coupled to Sepharose at a concentration of 4800 units of FVIII-LC: Ag per ml Sepharose, 0.2 ml of the immunosorbent was able to bind 900 Bethesda units from 100 ml hemophilia A inhibitor plasma. This opens the possibility to remove FVIII inhibitor antibodies from circulation by extracorporeal immunotherapy with FVIII-LC coupled to Sepharose.

Sign in / Sign up

Export Citation Format

Share Document