Poster Session II
Project Type
Poster
Project Funding and Affiliations
Center for Translational Medicine
Faculty Mentor’s Full Name
Blair DeBuysscher
Faculty Mentor’s Department
Health Department of Biomedical and Pharmaceutical Sciences
Abstract / Artist's Statement
Fentanyl overdose deaths are on the rise from both intentional use and accidental exposure. Our work focuses on identifying additional treatment options to Narcan, an opioid receptor antagonist which reverses overdose effects by receptor competition. One alternative method, that has been approved by the FDA, is the development of monoclonal antibodies (mAbs). This therapeutic method works by antibodies binding fentanyl in the bloodstream and preventing it from crossing the blood-brain barrier. Our group has begun identifying mAbs by vaccinating mice with a fentanyl hapten (F1) vaccine adjuvanted with a TLR 7/8 agonist. Vaccination leads to the production of antibodies that are specific to the fentanyl hapten. After vaccination, we isolated F1 specific B cells from lymph nodes of the mice and amplified the cDNA of the cells. Single cell sequencing of these cells allows us to identify, characterize and quantify the potential mAb candidates. To test therapeutic effects of these antibodies, we have midwifed plasmids to contain heavy and light chain constant fragments, used in combination to produce a fully functional antibody. We then cloned the variable regions from our selected sequences into each plasmid and transfected these into human 293F cells. After secretion and purification, antibodies are evaluated for F1 binding by Biolayer Interferometry. Samples with the highest measured affinity/avidity will be tested for efficacy against fentanyl using a challenge study in mice.
Category
Life Sciences
Combating Overdose: Production and Analysis of Fentanyl Specific Antibodies
UC South Ballroom
Fentanyl overdose deaths are on the rise from both intentional use and accidental exposure. Our work focuses on identifying additional treatment options to Narcan, an opioid receptor antagonist which reverses overdose effects by receptor competition. One alternative method, that has been approved by the FDA, is the development of monoclonal antibodies (mAbs). This therapeutic method works by antibodies binding fentanyl in the bloodstream and preventing it from crossing the blood-brain barrier. Our group has begun identifying mAbs by vaccinating mice with a fentanyl hapten (F1) vaccine adjuvanted with a TLR 7/8 agonist. Vaccination leads to the production of antibodies that are specific to the fentanyl hapten. After vaccination, we isolated F1 specific B cells from lymph nodes of the mice and amplified the cDNA of the cells. Single cell sequencing of these cells allows us to identify, characterize and quantify the potential mAb candidates. To test therapeutic effects of these antibodies, we have midwifed plasmids to contain heavy and light chain constant fragments, used in combination to produce a fully functional antibody. We then cloned the variable regions from our selected sequences into each plasmid and transfected these into human 293F cells. After secretion and purification, antibodies are evaluated for F1 binding by Biolayer Interferometry. Samples with the highest measured affinity/avidity will be tested for efficacy against fentanyl using a challenge study in mice.