3 results about "Fab Fragments" patented technology

The present invention provides bispecific antibodies of a new antibody format, and which comprise one or more Fab fragments and an immunoglobulin molecule. The invention further provides compositions of said bispecific antibodies, as well as methods and uses of the same. The invention further provides a method of generating bispecific antibodies of the new format.

This application relates to site-specific conjugation of antibody lysine residues using solid phase immobilized microbial transglutaminase MTG and MTG in solution. Site-specific modification of proteins using microbial transglutaminase (MTG) is a powerful and versatile strategy for controlled modification of proteins under physiological conditions. We present evidence that solid phase microbead immobilization can be used to site-specifically and efficiently link different functional molecules important for further downstream applications to therapeutically relevant proteins, including scFV, Fab fragments and antibodies. We demonstrate that MTG remains firmly immobilized and there is no detectable column bleed and the enzyme activity is maintained during continuous operation, which allows for convenient recycling of the enzyme, thus outperforming solution phase MTG conjugation. Furthermore, it is shown that immobilized MTG exhibits an enhanced selectivity for certain residues in the presence of several reactive residues, all of which are targeted if conjugation is performed in solution. Site-specific lysine conjugation of antibodies using immobilized and solution MTG with a highly efficient glutamine-containing peptide is also reported. In addition, the generation of bis-site-specific conjugated IgGl with site-specific conjugation of both glutamine and lysine residues of an IgGl antibody using immobilized and solution MTG is reported. Site-specific glutamine conjugation with small peptides containing lysine residues and functional moieties is also described.

A method is provided for determining the relative unpaired glycan content of an IgG antibody composition. In an exemplary embodiment, the method comprises (b) treating the IgG antibody composition with two enzymes to form a mixture of Fab fragments and Fc fragments, wherein one enzyme cleaves the antibody heavy chain at the N-terminal site of the hinge region disulfide bond and the other enzyme cleaves the β1,4 bond between core GlcNAc residues to form Fc fragments, each containing a pair of coreglycan structures; (b) separating the Fab fragments from the Fc fragments; and (c) quantifying the abundance of (i) paired non-fucosylated Fc fragments, (ii) unpaired non-fucosylated Fc fragments, (iii) paired high-mannose Fc fragments, and / or (iv) unpaired high-mannose Fc fragments to determine the relative unpaired non-fucosylated (AF) glycan content and the relative unpaired high-mannose (HM) glycan content.