Functional properties of antibody insulin-like growth factor fusion proteins.
AuthorsShin SU, Friden P, Moran M, Morrison SL
JournalJ Biol Chem
PubMed ID8106473
Genetic engineering and expression techniques have been used to produce antibody growth factor fusion proteins. Insulin-like growth factors (IGFs) 1 and 2 have been fused to mouse-human chimeric IgG3 at the end of CH1, immediately after the hinge, and at the end of CH3. Fusion heavy chains of the expected ... More
Cryoprecipitation properties of a high-affinity monoclonal IgM anti-fluorescyl antibody.
AuthorsDombrink-Kurtzman MA, Voss EW
JournalMol Immunol
PubMed ID3237215
'A high affinity (Ka = 2-3 x 10(10) M-1) murine monoclonal anti-fluorescein IgM antibody (18-2-3) exhibiting low temp. insolubility in the absence of bound ligand has served as a model to study cryoprecipitation. Insolubility of 18-2-3 at low temp. (4 degrees C) had been shown to be reversible at higher ... More
Binding of fluorescent and spin-labeled C-terminal hirudin analogs to thrombin.
AuthorsSankarapandi S, Woodford JK, Krstenansky JL, Berliner LJ
JournalJ Med Chem
PubMed ID7966143
'Synthetic peptides based on the sequence of the negatively charged carboxyl tail of hirudin exhibit anticoagulant activity. Several antithrombin agents are being developed by chemical and structural optimization of these "hirupeptides". The present work demonstrates the design and use of novel spin-labeled and fluorescent-labeled C-terminal hirudin analogs to study the ... More
Characterization of fluorescent 2-dimethylaminonaphthalene-5-sulfonyl-immunoglobulin G conjugates for application in fluorescence polarization studies.
AuthorsHerron JN, Voss EW
JournalJ Biochem Biophys Methods
PubMed ID7276421
'Preparations of normal rabbit IgG antibody (NgG) and purified rabbit IgG antibody of anti-2,4-dinitrophenyl (Anti-Dnp) specificity were covalently labeled with the fluorescent probe 2-dimethylaminonaphthalene-5-sulfonyl chloride (2,5-Dns-Cl). 2,5-Dns-immunoglobulin G preparations were shown to have average fluorescence lifetimes of 29--32 ns by cross-correlated phase and modulation fluorescence lifetime measurements. Excitation maxima were ... More
Fibrin-stabilizing factor inhibitors. IX. An inhibitor assay method based on SDS-disc gel electrophoresis.
Surface density determination in membranes by fluorescence energy transfer.
AuthorsFung BK, Stryer L
JournalBiochemistry
PubMed ID728398
Inter-active-site distance and solution dynamics of a bivalent-bispecific single-chain antibody molecule.
AuthorsMallender WD, Ferreira ST, Voss EW, Coelho-Sampaio T
JournalBiochemistry
PubMed ID8060979
The solution dynamics of a bivalent bispecific single-chain antibody (BiSCA) specific against fluorescein (Fl) and single-stranded DNA (ssDNA) were investigated. Fluorescence resonance energy transfer (FRET) studies were performed in order to estimate the average distances, R, between the anti-Fl and the anti-ssDNA active sites. In separate experiments, either 2-(dimethylamino)naphthalene-5-sulfonyl chloride ... More
Anti-metatype antibodies stabilize the fluorescein single-chain antibody 4-4-20 complex against dissociation by hydrostatic pressure.
AuthorsCoelho-Sampaio T, Voss EW
JournalJ Biol Chem
PubMed ID8132540
Hydrostatic pressure was used to promote dissociation of fluorescein (Fl) from single-chain antibody 4-4-20 (SCA 4-4-20). Fl fluorescence intensity was quenched by 97% upon binding to SCA 4-4-20. Increasing pressure to 2.4 kbar enhanced Fl fluorescence from the remaining 3% to 14-17%. The capacity of anti-metatype antibodies (anti-Met), which specifically ... More
Zymogen/enzyme discrimination using peptide chloromethyl ketones.
AuthorsWilliams EB, Krishnaswamy S, Mann KG
JournalJ Biol Chem
PubMed ID2708377
Glutamylglycinylarginyl chloromethyl ketone, tyrosylglycinylarginyl chloromethyl ketone, and phenylalanylprolylarginyl chloromethyl ketone have been labeled at their amino termini using fluorescein, rhodamine-X, lissamine-rhodamine, pyrene, and the 1,5-, 2,5-, and 2,6-dimethylaminonaphthalene-1-sulfonyl moieties. These peptidyl chloromethyl ketones have also been modified by incorporation of biotin and epsilon-amino caproyl biotin. The ability of these various ... More
A correlation between differential structural features and the degree of endopeptidase activity of type A botulinum neurotoxin in aqueous solution.
AuthorsCai S, Singh BR
JournalBiochemistry
PubMed ID11294637
Botulinum neurotoxin type A is one of the most toxic substances known to man (LD(50) for mouse 0.1 ng/kg). It is also an effective therapeutic drug against involuntary muscle disorders and for pain management. BoNT/A is a Zn(2+) endopeptidase which selectively cleaves SNAP-25 (synaptosomal-associated protein of 25 kDa), a critical ... More