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Design of antibody‐blocking monobodies: A novel therapeutic approach for autoimmune kidney disease
Author(s) -
DIPAK GALANI Aryan,
Stoddard Shana Victoria
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.472.4
Subject(s) - epitope , antigen , homology modeling , autoimmune disease , autoantibody , antibody , epitope mapping , autoimmunity , membranous nephropathy , in silico , immune system , chemistry , computational biology , protein engineering , immunology , biology , biochemistry , kidney , glomerulonephritis , genetics , enzyme , gene
Autoimmune diseases arise when antibodies erroneously attack healthy cells. Primary membranous nephropathy (PMN) is a kidney specific autoimmune disease affecting 10–12 million individuals worldwide. Currently non‐specific immunosuppressants which weaken the body's entire immune system are used to treat many autoimmune disorders. These non‐specific treatments decrease the patient's ability to fend off basic infections. Thus, there is a need for more targeted approaches to autoimmune diseases therapies. Therefore the goal of this project is the development of an antigen specific approach for autoimmune disease, through the design of protein monobodies capable of preventing autoantibody binding to the antigen. The PMN antigen, thrombospondin type‐1 domain‐containing 7a (THSD7A), was used as a target for cap development. Epitope sites were identified using an in silico homology model for the THSD7A antigen. The work here focuses on targeting two of the 17 extracellular domains which were identified to contain epitope sites, (specifically domains 9 and 10). In silico mutagenesis to the 3QHT monobody template was performed using Chimera and Phyre 2 programs. Interface scores measured in Rosetta energy units (REU) were improved from −3.129 (3QHT template) to −3.927 in the 3QHT‐015‐AG cap. Data shows that introduction of hydrophobic residues enhanced binding. Introduction of a Phe residue, creating the 3QHT‐023‐AG mutant, permitted a cation‐pi interaction to domain 9 of THSD7A resulting in an increased binding score of −6.617 REU. Currently protein expression and purification is underway to allow for ELISA assays and further evaluation of these epitope caps binding to domain 9 of the THSD7A antigen. Through the development of antigen specific blocking monobodies, successful caps could provide a much safer and accurate treatment for autoimmune diseases. Support or Funding Information Faculty Development Endowment Grant, Rhodes College This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .