Disulfide bonds monoclonal antibodies

S-Fl Jung, I Pastan, B Lee. Design of interchain disulfide bonds m the framework region of the Fv fragment of the monoclonal antibody B3. Pi otems 19 35-47, 1994. 

In a comparative study of disulfide reducing agents, it was determined that use of the relatively strong reductants DTT and TCEP required only 3.25 and 2.75 mole equivalents per mole equivalent of antibody molecule to achieve the reduction of two interchain disulfide bonds between the heavy chains of a monoclonal IgG (Sun et al., 2005). This limited reduction strategy retains intact bispecific antibody molecules while providing discrete sites for conjugation to thiols. 

B. Packard and M. Edidin, Site-directed labeling of a monoclonal antibody Targeting to a disulfide bond, Biochemistry 25, 3548-3552 (1986). 

Hyde, G.E., Wilberding, J.A., Meyer, A.L., Campbell, E.R. Campbell, W.H. (1989). Monoclonal antibody-based immunoaffinity chromatography for purifying corn and squash NADH nitrate reductases. Evidence for an interchain disulfide bond in nitrate reductase. Plant Molecular Biology 13, 233-46. 

More recently, this model has undergone substantial modification. First, it became apparent from biosynthetic studies on cells in culture that there were two distinct B chains, B1 and B2 (Cooper et al., 1981), which were not well resolved in laminin prepared from the EHS tumor. Analysis of the stoichiometry of the chains produced in these systems suggested that the laminin molecule contained one A, one Bl, and one B2 chain (Fig. 9). Studies with a monoclonal antibody specific for the A chain indicated that the A chain formed part of the short arm as well as the long arm of laminin (Palm et al., 1985). The assembly of the molecule appears to proceed in discrete steps, with the initial assembly of a B1-B2 dimer linked by disulfide bonds to which an A chain is added (Morita et al., 1985 Peters et al., 1985). 

Elhott S, Chang D, Delorme E, Dunn C, Egrie J, Giffin J, Lorenzini T, Talbot C, Hesterberg L. Isolation and characterization of conformation sensitive antieryth-ropoietin monoclonal antibodies effect of disulfide bonds and carbohydrate on recombinant human erythropoietin structure. Blood 1996 87 2714-22. 

Brody, 1. (1997), Multistep dcnaiuration and hierarchy of disulfide bond cleavage of a monoclonal antibody. AiwJ. Biochem. 247,247-256. 

More productive chemical results, which stiU harness the destructive action of ultrasound on certain bonds, can be attained when sonication is applied to biological fluids (e.g. protein solutions) en route to bionanomaterials [15], A conspicuous example can be found in sonochemically-prepared protein microspheres, in which the interplay of mechanical effects (emulsification) and chemical effects (formatiOT of transient species) is noticeable. A protein emulsion is readily created at the interface between two immiscible liquid phases, while radicals generated by water sonolysis promote disulfide bond cross-linking between cysteine residues. Surface modifications, via conjugation with monoclonal antibodies or RGD-containing peptides, can also be carried out [102, 103]. The sonochemical preparation of chitosan microspheres also exploits the intermolecular cross-linking of imine bonds from the sugar precursor [104]. 

Monoclonal antibodies are inherently stable proteins. Being secretory products they are stabilized by disulfide bonds and not readily susceptible to damage by oxidation. In our experience, culture supernatants containing IgG antibodies remained active for at least 15 years at 4 °C when Hepes and sodium azide were... 

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