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Increasing the efficiency and productivity of mAb or AAV and lentiviral vector-based product titer determination is becoming more critical with the need to support multiple programs with limited resources. Limitations in current methods such as enzyme-linked immunosorbent assay (ELISA) or high-performance liquid chromatography (HPLC) such as low throughput, limited dynamic range, and long assay times create bottlenecks in critical workflows. In contrast, Gyrolab automated immunoassay platforms generate high-quality analytical data with high throughput using nanoliter volumes to enable more rapid and efficient product titer determinations.
Selection of stable, high-producing clones during cell line development (CLD) is subject to aggressive timelines. Hundreds of clones often require rapid assessment for mAb product titer analysis. The figure above shows the titer distribution of all clones in a screen for low culture titers at early stage CLD. IgG titers of post-transfection samples from 96-well plates were measured using Gyrolab xP platform, which processed 332 samples in four hours. Clones with high IgG titers were further expanded.
Gyrolab platforms offer key advantages for titer analysis in applications such as protein expression, CLD, and viral vector titer for lentiviral and AAV vector-based gene therapies. The robust, nanoliter-scale automated platform provides high throughput and rapid assay results to support data-driven decision making.
|Sample volume (µL)||10 -100 µL||4 µL (including dead volume)|
|Measurement range||0.02 - 5 mg/mL (usually 1 - 50 µg load); sample dilution can be avoided by adjusting sample injection volumes||0.04 - 6 mg/mL;
single 5x dilution for all samples
Intra (neat)≤ 3%
Inter (neat)≤ 3%
Intra (5x dilution) <4%
Inter (5x dilution) <11%
|Assay time including sample preparation
(e.g. 100 samples)
|13 – 16 hours||< 2 – 3 hours|
|Accuracy||88 – 119%||99 – 112%|
Gyrolab human IgG titer Kits for low titer and high titer with assay run times of about an hour provide rapid quantification of intact human IgG (IgG1, IgG2, IgG4) in cell culture supernatants during cell line development and production. The wide assay range of over 6 logs minimizes dilutions and repeats for analysis.
Quantification of viral vector titers requires precise, robust assay methods that ideally use a minimum sample volume and have rapid completion times. Gyrolab immunoassays are ideally suited for this purpose, requiring <10 µL of sample, sparing precious production batch sample volumes.
With Gyrolab kits for measurement of AAV total capsids or p24 physical lentiviral titer, assay development is eliminated. Measurement of total vector titer when combined with orthogonal analytical methods such as digital droplet PCR (ddPCR), quantitative PCR (qPCR), analytical ultracentrifugation (AUC), or size exclusion HPLC, empty/full titer ratios can be determined.
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