Engineered Protease-resistant Antibodies with Selectable Cell-killing Functions*
By M. Kinder, A. Greenplate, Katharine D. Grugan, Keri L. Soring, Katharine Heeringa, Stephen G McCarthy, G. Bannish, Meredith Perpetua, F. Lynch, R. Jordan and 2 more. 2013
41 citations recorded by the source corpus.
Background: Proteases can cleave human IgG1 antibodies, resulting in loss of cell-killing functions. Results: Mutation of the lower hinge of IgG1 confers protease resistance but disrupts Fc effector functions. Conclusion: Compensating mutations in the CH2 domain can selectively restore Fc effector functions on a protease-resistant backbone. Significance: Protease-resistant antibodies may be desirable for microenvironments with high protease content and/or when selected cell-killing functions are needed. Molecularly engineered antibodies with fit-for-purpose properties will differentiate next generation antibody therapeutics from traditional IgG1 scaffolds. One requirement for engineering the most appropriate properties for a particular therapeutic area is an understanding of the intricacies of the target microenvironment in which the antibody is expected to function. Our group and others have demonstrated that proteases secreted by invasive tumors and pathological microorganisms are capable of cleaving human IgG1, the most commonly adopted isotype among monoclonal antibody therapeutics. Specific cleavage in the lower hinge of IgG1 results in a loss of Fc-mediated cell-killing functions without a concomitant loss of antigen binding capability or circulating antibody half-life. Proteolytic cleavage in the hinge region by tumor-associated or microbial proteases is postulated as a means of evading host immune responses, and antibodies engineered with potent cell-killing functions that are also resistant to hinge proteolysis are of interest. Mutation of the lower hinge region of an IgG1 resulted in protease resistance but also resulted in a profound loss of Fc-mediated cell-killing functions. In the present study, we demonstrate that specific mutations of the CH2 domain in conjunction with lower hinge mutations can restore and sometimes enhance cell-killing functions while still retaining protease resistance. By identifying mutations that can restore either complement- or Fcγ receptor-mediated functions on a protease-resistant scaffold, we were able to generate a novel protease-resistant platform with selective cell-killing functionality.
Subfields: Immunology, Molecular Biology, Radiology, Nuclear Medicine and Imaging
How researchers see this paper
The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.
In Molecular Biology
In Radiology, Nuclear Medicine and Imaging
Citations in the corpus
- Antibody Structure and Function: The Basis for Engineering Therapeutics (2019, background): “N-linked glycosylation can be removed by using the mutation N297A on IgG1, IgG2, IgG3, or IgG4.”