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anna.moberg@ge.com  
#1 Posted : 14 September 2017 11:50:11(UTC)
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GE'anna.moberg@ge.com

Joined: 30/06/2017(UTC)
Posts: 3

Recently I came across a publication written by Vishal Kamat and Ashique Rafique from Regeneron Pharmaceuticals. The title of the publication is Extending the throughput of Biacore 4000 biosensor to accelerate kinetic analysis of antibody-antigen interaction. In this publication, Vishal Kamat and Ashique Rafique describes a very clever way of utilizing the parallel configuration and the hydrodynamic addressing possibilities of Biacore 4000 to develop three kinetics assays for formats not normally included in Biacore 4000. The first assay is a capture assay. The normal capture procedure of Biacore 4000 allows for capture of up to 8 different mAbs in parallel. By utilizing the potential of the instrument in a slightly different way, the authors of this paper managed to capture a total of 16 different mAbs (spots 1, 2, 4 and 5 in each flow cell). This was accomplished the following way: 8 different Mabs were captured on 16 spots immobilized with anti-human IgG (same mAb on spots 1 and 2 as well as on spots 4 and 5 in each flow cell). After that, a general injection was used to regenerate and remove the antibodies on the outer spots (1 and 5) of each flow cell. In the final step, 8 additional mAbs were captured on spots 1 and 5 in each flow cell, which makes a total of 16 different mAbs in parallel with spot 3 used as reference spot. Concentration series of four different antigens were then run over the surface according to standard procedure. The second assay was run in single-cycle kinetics format. For this assay, a total of four different antigen concentrations were injected over the mAb surface. The Sample command was used for the lowest concentration. For the additional concentrations the General injection command was applied. A dissociation time was set by utilizing the command Stabilization period after injection. After the last injection this time was set to 600 s in order to maximize the dissociation time. By adding an enhancement injection after the stabilization period it was possible to extend the dissociation phase with another 300 s. The third assay was a parallel kinetics assay. This format is currently only available for Biacore 8K. In this assay, the same mAb is captured across different flow cells followed by injections of different concentrations of antigen across the flow cells. A blank cycle using only buffer as the analyte was included for each mAb. Thus, a complete kinetics experiments is performed in only two cycles. Evaluation of the different assays was performed using Biacore 4000 Evaluation Software combined with either Scrubber or BIAEvaluation software. The authors found that the three assays provided reproducible and reliable kinetic data and led to a better utilization of Biacore 4000 and an increased throughput. By implementing the new assays the mAb characterization workflow can be accelerated which will result in a more rapid selection of lead candidates. Very nice data and a very nice way of utilizing the possibilities of the instrument! Anna Moberg, GEHC BioSciences Reference: Extending the throughput of Biacore 4000 biosensor to accelerate kinetic analysis of antibody-antigen interaction, Vishal Kamat and Ashique Rafique, Analytical Biochemistry 530 (2017) 75-86.
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