کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5131685 | 1491317 | 2017 | 12 صفحه PDF | دانلود رایگان |
- Three new capture kinetic assays were developed to increase throughput - 16-mAb capture kinetic, single cycle kinetic (SCK) and parallel kinetic (PK).
- All three capture kinetic assays generate reliable and reproducible binding kinetic parameters.
- The 16-mAb capture kinetic assay reduces experiment time and analyte consumption by 35% and 50%, respectively.
- Both SCK and PK reduce assay development time by avoiding surface regeneration, which further extends the throughput of Biacore 4000.
The surface plasmon resonance (SPR) biosensors are being routinely used in different stages of drug discovery and development. However, the lack of high throughput SPR biosensors continues to be a primary bottleneck for the rapid kinetic screening of large panels of monoclonal antibodies (mAbs). To further increase the throughput of the Biacore 4000 biosensor, we have developed three kinetic screening assays to characterize mAb-antigen interactions - (i) 16-mAb capture kinetic, (ii) single cycle kinetic (SCK), and (iii) parallel kinetic (PK). The performance of all three kinetic assays was evaluated by characterizing the binding of kinetically diverse human mAbs to four antigens with molecular weights of 14kD, 29kD, 38kD, and 48kD and binding affinities ranging from 130pM to 200Â nM. The binding rate constants measured using all three kinetic assays were reproducible across multiple experiments and correlated with the values generated using the conventional 8-mAb capture kinetic assay on the Biacore 4000 (R2Â >Â 0.94). Moreover, the 16-mAb capture assay decreased experiment time and analyte consumption by 35% and 50%, respectively. This work illustrates the significance of the 16-mAb capture kinetic, SCK, and PK assays to increase the throughput of Biacore 4000 and to support rapid kinetic screening of mAbs.
Journal: Analytical Biochemistry - Volume 530, 1 August 2017, Pages 75-86