Engineering pH dependent camelid antibodies with aspartic acid and glutamic acid

Advisor(s)

James R. Horn, Northern Illinois University

Hyeyoung Eom, Northern Illinois University

Location

Room A123-2

Start Date

26-4-2019 11:05 AM

End Date

26-4-2019 11:20 AM

Abstract

Protein engineering is growing as an important subject as new discoveries in this area open new doors for application of different proteins. Camelid antibodies specifically are an area of interest because they lack the light chain and only possess the heavy chain that allow for unique applications. This heavy-chain only design increases stability and specificity in bonding interactions. Previously, camelid antibody proteins were engineered by the Horn Lab that had high bonding affinities at high pHs. Using isothermal titration calorimetry, we studied the bonding affinities at pH 4.0 in several mutations that allowed for high binding at low pH. By mutating certain amino acids in the homodimer bonding interface to aspartic acid or glutamic acid, we were able to engineer proteins with different structures to examine the efficacy and the affinity of the bonding at low pH. Further research is needed to determine the most effective mutation and to study the effect these mutations have on the structure.

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Apr 26th, 11:05 AM Apr 26th, 11:20 AM

Engineering pH dependent camelid antibodies with aspartic acid and glutamic acid

Room A123-2

Protein engineering is growing as an important subject as new discoveries in this area open new doors for application of different proteins. Camelid antibodies specifically are an area of interest because they lack the light chain and only possess the heavy chain that allow for unique applications. This heavy-chain only design increases stability and specificity in bonding interactions. Previously, camelid antibody proteins were engineered by the Horn Lab that had high bonding affinities at high pHs. Using isothermal titration calorimetry, we studied the bonding affinities at pH 4.0 in several mutations that allowed for high binding at low pH. By mutating certain amino acids in the homodimer bonding interface to aspartic acid or glutamic acid, we were able to engineer proteins with different structures to examine the efficacy and the affinity of the bonding at low pH. Further research is needed to determine the most effective mutation and to study the effect these mutations have on the structure.