The present invention provides a method for manipulating the fucosylated glycan content on a recombinant protein.

The present invention provides a method for manipulating the fucosylated glycan content on a recombinant protein.

Disclosed is an isolated monoclonal antibody against a human Transforming Growth Factor-β (TGF-β) Latency Associated Protein (LAP) degradate, the isolated monoclonal antibody being capable of recognizing an integrin binding site in the human TGF-β LAP degradate.

Through a combination of in vitro and in vivo approaches, the inventors show that human lgG2 (h2) delivers unique FcγR-independent agonistic activity to anti-CD40 antibodies and to antibodies specific to other immunostimulatory receptors, including 4-1BB and CD28. Investigation of an anti-human CD40 mAb, LOB7.4, revealed that the unique activity of h2 was dependent upon the precise arrangement of hinge and CH1 disulfide bonds. Chemical ‘shuffling’ or mutagenesis to ‘lock’ LOB7.4 into either a more flexible ‘h2A’ or more compact ‘h2B’ conformation endowed antagonistic and agonistic properties, respectively. Engineering of h2 in this way allows development of reagents with either immunostimulatory or immunosuppressive characteristics, with direct implication for the design of therapeutic mAb agents and fusion proteins.

Through a combination of in vitro and in vivo approaches, the inventors show that human IgG2 (h2) delivers unique FcγR-independent agonistic activity to anti-CD40 antibodies and to antibodies specific to other immunostimulatory receptors, including 4-1BB and CD28. Investigation of an anti-human CD40 mAb, LOB7.4, revealed that the unique activity of h2 was dependent upon the precise arrangement of hinge and CH1 disulfide bonds. Chemical ‘shuffling’ or mutagenesis to ‘lock’ LOB7.4 into either a more flexible ‘h2A’ or more compact ‘h2B’ conformation endowed antagonistic and agonistic properties, respectively. Engineering of h2 in this way allows development of reagents with either immunostimulatory or immunosuppressive characteristics, with direct implication for the design of therapeutic mAb agents and fusion proteins.

The present invention provides a method for manipulating the fucosylated glycan content on a recombinant protein.

Disclosed is an isolated monoclonal antibody against a human TGF- LAP degradate, the isolated monoclonal being capable of recognizing an integrin binding site in the human TGF- LAP degradate.

The present invention provides a method for manipulating the fucosylated glycan content on a recombinant protein.

The present invention provides a method for manipulating the fucosylated glycan content on a recombinant protein.

A polypeptide comprising a chimeric antigen receptor (CAR) comprising (i) an extracellular domain capable of binding to a first antigen, (ii) a transmembrane domain, and (iii) an intracellular domain; and a domain capable of binding to a second antigen expressed on the surface of a cell that can interact with a T cell, wherein the CAR and the domain are fused by a peptide linker.