Monoclonal antibodies are of particular interest in basic biomedical research and for the diagnosis and treatment of various diseases. Screening, profiling, and production of antibodies involve tedious and time-consuming procedures. Bio-Rad now offers the ProteOn XPR36 protein interaction array system, a label-free, multiplex system for rapid screening and comprehensive kinetic profiling of antibody interactions.
The ProteOn XPR36 instrument is based on surface plasmon resonance (SPR) technology and meets the challenges of high-throughput processing by bringing a novel one-shot kinetics approach to the study of protein interactions. The system's parallel and real-time sample processing capabilities enable simultaneous acquisition of comprehensive data for 36 molecular interactions — without the need for regeneration.
Protocol for Selection and Ranking of 250 Mouse Monoclonal Antibody Supernatants against Human Interleukin 12 (IL-12)
Anti-mouse IgG is immobilized covalently on six microfluidic channels using a ProteOn GLM sensor chip (see Figure 1 for protocol workflow).
Six different unpurified mouse anti-human IL-12 supernatant solutions are injected through six orthogonal channels in parallel. Specific mouse antibodies are captured by the immobilized anti-mouse IgG capture agent.
Five concentrations of purified IL-12 antigen are applied simultaneously in the orientation of the original anti-mouse IgG capture antibodies. Running buffer is injected in the sixth channel as a double reference to correct for any decay of the captured anti-IL-12 from the anti-mouse IgG. Thirty-six sensorgrams (six sets of six) are generated simultaneously, providing the association and dissociation kinetics of all six antibodies.
IL-12 antigen-antibody pairs are removed using diluted phosphoric acid, allowing capture and analysis of a new panel of 6 hybridoma supernatants.
Repeating the cycle 42 times on the same chip allows determination of a comprehensive kinetic profile for all 250 supernatants in less than 13 hours, enabling rapid and high-throughput screening for high-affinity cell lines (see Figure 2).
The multiplexing and one-shot kinetics capabilities of the Bio-Rad ProteOn XPR36 system provide a powerful approach for the rapid screening of large numbers of hybridoma supernatants, a previously time-consuming and laborious component in the area of antibody research.
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