Direct sequencing of serum antibodies after infection

Project Summary Monoclonal antibodies mAbs are a well validated drug platform with exquisite specificity diversity and potency They offer the lowest risk class of drug for development through licensure and offer great potential for addressing emerging and re emerging infectious diseases To be prepared as threats like Ebola EBOV Marburg MARV and Zika viruses continue to emerge or re emerge rapid discovery capabilities are a critical element One of the best current methods for discovery of potent human mAbs is isolation of peripheral B cells from survivors sero positive individuals for single cell sequencing or hybridoma generation However peripheral B cells are not always easy to obtain and only represent a small percentage of the total B cell population across all bodily tissues We have developed an antibody discovery technology in which only serum antibodies are required i e mAb sequences against a given antigen can be sequenced de novo from polyclonal antibody pAb pools without the need for sequencing of genetic material We propose to further refine this novel proteomic approach and apply it towards discovery of new antibodies in serum obtained from EBOV survivors Here IgGs from survivor plasma will be purified by filovirus glycoprotein antigen specificity and sequenced independently of B cells Given our preliminary data these efforts may yield mAbs that are cross reactive to ZEBOV SUDV BDBV and possibly MARV Viable mAbs will ultimately be developed to become pan EBOV ZEBOV SUDV BDBV and or pan filovirus EBOV MARV therapeutic products But perhaps the most significant contribution of this work will be to further develop our rapid antibody discovery approach which may impact drug discovery in nearly all sectors of the mAb industry including infectious disease Narrative The focus of this proposal is to optimize an antibody discovery technology that utilizes direct sequencing of serum antibodies This discovery technology will be applied to serum from survivors of Ebola virus disease There is a clear unmet need for improved and rapid antibody discovery that can identify cross reactive high affinity mAbs against a variety of emerging viruses The identified antibodies may offer improved therapeutic options since they are derived from human survivors