Site-directed labeling of a monoclonal antibody: targeting to a disulfide bond.

@article{Packard1986SitedirectedLO,
  title={Site-directed labeling of a monoclonal antibody: targeting to a disulfide bond.},
  author={Beverly Z. Packard and Michael A Edidin and A Komoriya},
  journal={Biochemistry},
  year={1986},
  volume={25 12},
  pages={
          3548-52
        }
}
We have designed and synthesized crabescein, the first member of a class of fluorescent labels that add across disulfide bonds. Crabescein is a fluorescein derivative that reports the rotational correlation time of the immunoglobulin G (IgG) segment to which it is covalently bound. Chemical analysis of the IgG labeled with crabescein indicates that the fluorophore is inserted into the third disulfide bond (cysteine-229 of mouse IgG2a) in the hinge region. The rotational correlation time of this… Expand
Disulilde Bond-targeted Radiolabeling: Tumor Specificity of a Streptavidin- biotinylated Monoclonal Antibody Complex1
A site-specific labeling method was developed in which sulfhydryl groups of a murine IgG2a anti-ovarian monoclonal antibody, 5G6.4, were biotinylated with /V-iodoacetyl-yV'-biotinylhexylenediamineExpand
Site-directed chemical modification and cross-linking of a monoclonal antibody using equilibrium transfer alkylating cross-link reagents.
TLDR
A new, more reactive group of protein cross-linkers in the class of equilibrium transfer alkylating cross- link (ETAC) reagents has been synthesized and had a biodistribution in CD1 mice similar to that of the 111In bis cyclic anhydride DTPA labeled antibody. Expand
Sulfhydryl site-specific cross-linking and labeling of monoclonal antibodies by a fluorescent equilibrium transfer alkylation cross-link reagent.
TLDR
The present study illustrates the potential applications of labelable ETAC reagents as thiol-specific probes for a wide variety of immunological studies. Expand
Preparation, characterization, and in vivo biodistribution properties of synthetically cross-linked multivalent antitumor antibody fragments.
Two new antibody forms of the general structure F(ab')n (n = 3 or 4) were prepared and tested in vivo as part of an ongoing search for antibody candidates with improved biodistribution properties forExpand
Site-specific radiolabeling of monoclonal antibodies with biotin/streptavidin.
  • R. D. del Rosario, R. Wahl
  • Chemistry, Medicine
  • International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology
  • 1989
TLDR
Sulfhydryl groups of a murine IgG2a monoclonal antibody were site-specifically biotinylated with N -iodo-acetyl- N′ -biotinylhexylenediamine following partial reduction of disulfide bonds and in vivo localization of the radioantibody complex to xenografts of ovarian carcinoma was achieved. Expand
Chemical modification of rabbit IgG with N-dansylaziridine. Investigation of the properties of dansylated antibodies.
TLDR
The result indicates that interaction of antibodies with antigen is accompanied by the conformational changes in the IgG hinge region, and labels of IgG with DAZ does not alter its conformation, hydrodynamic behavior, nor its antigen binding properties. Expand
Effect of chemical modification strategy on the characteristics of copper-67-Labeled immunoconjugates, Part I: Immunoreactivity
AbstractThe synthetic porphyrins N-benzyl-5,10,15,20-tetrakis(4-carboxyphenyl)porphine (N-bzHTCPP) and N-4-nitrobenzyl-5-(4-carboxyphenyl)-10,15,20-tris(4-sulfophenyl)porphine (NbzHCS3P) representExpand
The role of thiols and disulfides on protein stability.
TLDR
The effect of introducing a disulfide bond to improve physical stability of proteins and the mechanisms of degradation of disulfides were discussed and the qualitative/quantitative methods to determine the presence of thiol in the Cys residue and various methods to derivatize thiol group for improving protein stability were illustrated. Expand
Comparison of four methods for the biofunctionalization of gold nanorods by the introduction of sulfhydryl groups to antibodies
TLDR
The introduction of a thiol group to antibodies by using Traut’s reagent, DTT, and PEG6-CONHNH2 allowed for direct immobilization onto the GNR surface, improved the efficacy of functionalized GNRs, and increased the sensitivity in response to target detection as a biosensor. Expand
The role of thiols and disulfides in protein chemical and physical stability
The use of proteins for therapeutic applications has increased dramatically in the last several decades. There has been a tremendous increase in the number of approved drugs derived from recombinantExpand
...
1
2
...

References

SHOWING 1-10 OF 21 REFERENCES
Segmental flexibility in an antibody molecule.
TLDR
The striking finding is that the antibody molecule displays a discrete mode of flexibility in the nanosecond time range, which exemplifies a structural design which is probably used in other large proteins and in organized macromolecular assemblies. Expand
Segmental flexibility of immunoglobulin G antibody molecules in solution: a new interpretation.
TLDR
The flexibility of native and mildly reduced anti-5-(dimethylamino)naphthalene-1-sulfonyl (anti-dansyl) antibody was reexamined by nanosecond fluorescence spectroscopy using deconvolution and lamp-shift corrections and indicates that the long correlation time primarily represents motions of the Fab segments and not global tumbling of the entire molecule. Expand
Rotational dynamics of immunoglobulin G antibodies anchored in protein A soluble complexes.
TLDR
The rotational dynamics of rabbit IgG anti-dansyl antibodies anchored in staphylococcal protein A (SpA) soluble complexes were studied by both steady-state and nanosecond fluorescence spectroscopy to provide rather striking evidence that phi L mainly represents flexible motions of the Fab segments and not global tumbling. Expand
The effects of cleavage of the inter-chain disulphide bonds of rabbit IgG on its ability to bind C1q.
TLDR
The ability of the IgG to form insoluble immune complexes was partially compromised when iodoacetamide was used to block the disulphide bonds, but was unimpaired when N-( iodoacetylaminoethyl )-8-naphthylamine-1-sulphonic acid was used. Expand
Nanosecond fluorescence spectroscopy of pyrenebutyrate anti-pyrene antibody complexes.
TLDR
The present investigation confirmed the presence of Fab segmental fexibility in immunoglobulin G molecules specific for a hapten different from DNS-lysine and obtained a value of about 135 ns for the longer rotational correlation time which probably represents global rotation of the entire molecule. Expand
A kinetic study in vitro of the reoxidation of interchain disulfide bonds in a human immunoglobulin IgGLk. Correlation between sulfhydryl disappearance and intermediates in covalent assembly of H2L2.
TLDR
It is concluded that the in vitro reoxidation of Fro is other than random; involved a pathway of pathways with HL, H2, and H2L precursors; and involves at least some kinetic cooperativity in bond formation, since no model bases solely on independent bond formation adequately accounts for the results. Expand
Some theoretical considerations regarding the effects of steric hindrance and intrinsic global coupling on the flexibility of Fc-anchored immunoglobulins.
TLDR
Estimates of the anisotropy contributions from flexible and global motions of the IgG-SpA complexes are determined by contrasting theoretical and measured decays and it is evident that the rabbit IgG anti-dansyl antibodies do not have much intrinsic global coupling, but rather they are highly flexible. Expand
The disulphide bonds of the heavy chain of rabbit immunoglobulin G.
TLDR
The cystine-containing peptides from enzymic digests of whole immunoglobulin G and Fc fraction were studied by several techniques and the results obtained enable us to put forward a scheme of the arrangement of the inter- and intra-chain disulphide bonds. Expand
Intracellular dye heterogeneity determined by fluorescence lifetimes.
The cellular localization of a fluorescent probe molecule depends on both the chemical structure of the dye and the cellular environment. To study the number and types of environments in anExpand
Conversion of incomplete antibodies to direct agglutinins by mild reduction: evidence for segmental flexibility within the Fc fragment of immunoglobulin G.
TLDR
Results suggest that mild reduction of the antibody imparts sufficient freedom to permit bridging between cells and are interpreted as evidence that the interheavy-chain disulfide bonds restrict segmental flexibility within the Fc fragment of IgG. Expand
...
1
2
3
...