Determination of mAb binding kinetics (ka & kd) and affinity (KD) are essential to optimum therapeutic candidate selection, with affinity ranking by association and/or dissociation constant commonly performed as a primary screening step. While SPR is the gold standard in kinetic characterization of therapeutic mAbs, other techniques are commonly used as a primary screening tool due to the low throughput, high sample requirements and limited data analysis capability of current SPR instruments.
Wasatch Microfluidics’ CFM/SPRi Array Platform changes the paradigm, delivering high throughput First-Pass Kinetics™ mAb screening:
- High throughput: Kinetically screen and rank 100’s of mAbs per day
- Low sample usage: Only ~100µl of each interactant required
- Ease-of-use: Simplified assay development using our unique Condition Scouting Tool
- Rapid data analysis: Dedicated software tools enable large data-sets to be rapidly fitted and visualized
- Detailed data mining: Interrogate mAb panels via interactive on-rate, off-rate, affinity and iso-affinity plots
- Data integration: Combine binning and kinetics screening data for un-paralleled mAb evaluation
Go beyond simple endpoint affinity or off-rate ranking and generate more granular data sets using full kinetics screening
Antibodies of markedly different association and dissociation kinetics can appear nearly identical when considering only endpoint affinity. Similarly, off-rate ranking alone can fail to meaningfully differentiate large groups of candidates. Combining measures of association and dissociation kinetics in a single experiment yields more thoughtful data sets that can be rapidly meshed with other criteria for candidate ranking. First-Pass Kinetics ™ facilitates robust selection of candidate antibodies with binding profiles better aligned to specific therapeutic objectives.
Two Strategies for First Pass Kinetics™ Screening:
First: Couple antibodies directly to the sensor chip using the CFM and probe with kinetic titration of analyte in parallel on the MX96
- Utilize with or without regeneration between analyte injections
- Extremely low antibody requirements
Second: Capture up to 96 distinct antibodies or a subset under multiple conditions with the CFM and perform kinetics screening in parallel on the MX96
- No regeneration between analyte injections; ideal for regeneration-sensitive antibodies
- Readily capture from crude supernatants
- Surface can be stripped at end of experiment and a new antibody set captured
Both strategies afford a full kinetics screen of up to 96 unique mAbs in a single day
This white paper describes the determination of the kinetic rate and equilibrium affinity constants of Fc – VH interactions as performed in the labs of Crescendo Biologics, Cambridge, UK.