무표지 생체분자 상호작용 분석을 위한 바이오층 간섭계

과학 연구의 획기적인 발전을 20년간 지원

수천 번 입증된 – 진정한 Octet BLI는 7,000개 이상의 동료 검토 출판물을 통해 다양한 연구 영역과 어플리케이션을 선도하는 기술입니다. Octet 출판물 데이터베이스는 실시간 무표지 생체분자 상호작용 분석을 위한 Octet 제품 사용을 참조 및/또는 인용하는 과학 출판물을 쉽게 검색할 수 있는 방법을 제공합니다.

Baseline Stability, Low Noise, and Low Baseline Drift are Key for High-Quality Data

The Octet R Series offers an advanced, fluidics-free approach to bimolecular analysis, with a wide variety of off-the-shelf biosensors for rapid binding kinetics and quantitation analysis. These systems utilize Sartorius’ genuine label-free Biolayer Interferometry (BLI) technology, enabling direct detection of specific proteins and other biomolecules — even in complex mixtures like crude cell culture supernatants and lysates.

The Octet BLI systems monitor binding events in real time to calculate on rates (), off rates () and affinity constants (). The superior sensitivity enables measurement of low molecular weight molecules down to 150 Da. The following example shows the kinetic interaction analysis of a small molecule.

The kinetic analysis of between a ligand, biotinylated-carbonic-anhydrase, and the small molecule analyte with Super Streptavidin (SSA) Biosensors at 1000 rpm and 25°C on an Octet R8 system. Working solutions were prepared in serial dilutions of 0.1, 0.3, 1.0. 3.0, 10.0 µM (n=4), respectively. Data was processed and curve fitted using a 1:1 binding model. 

Slow off-rate measurements show minimal changes in signals in the dissociation step requiring a combination of stable baselines and low baseline drifts for accurate measurement of the dissociation phase.

In addition, capturing measurable dissociation of the interaction may take hours of (> 2 hr data acquisition) specifically at the dissociation phase of higher affinity interactions that exhibit very slow off rates. Therefore, measuring longer dissociation times is also critical for measuring high-affinity interactions.

The Octet R8 system simultaneously enables measurements of up to 8 samples in a single unattended run. Combined with a unique microplate cover that keeps samples intact by minimizing evaporation, it allows scientists to collect accurate measurements with evaporation control for up to 12 hours.

The measurement of one high-affinity interaction measured in triplicates in a long dissociation experiment on the Octet R8 system is demonstrated below. Full kinetic characterization of an antibody-antigen interaction was run three times with a total run time of 11 hours.

Figures 2 and 3 provide two examples of long dissociation measurement over several hours to allow the detection of very slow off-rates.

Not Just GxP Compliance Tools — Rely on a Comprehensive Toolbox

From antibody quantitation, ligand binding characterization to potency assays, Sartorius offers a wide range of GxP compliance tools for developing validated assay methods on the genuine Octet BLI platform.

The United States Pharmacopeia (USP) has recently released a new general chapter <1108> which for the first time cites the use of biolayer interferometry in key ligand binding applications that provide more precise kinetic data and that can measure a relatively wide range of binding affinities. 

Sartorius Offers c 21 CFR Part 11 Software and a Full Line of GxP Products and Services as Part of the Octet GxP Package Including Our Biosensor Validation Support Services

This service enables the Octet BLI users to sample multiple lots of a biosensor during assay qualification and validation and reserve a well-characterized lot for purchase. Hence, customers can order up to five different biosensor lots, up to 20 trays from each lot for evaluation, and reserve up to 40 additional trays for future purchase.

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Approved Drugs Utilizing Octet BLI Data in Their Regulatory Approval Process

Octet BLI assays can help identify potential safety concerns and improve the accuracy of drug evaluations. Here are several examples of companies, including Pfizer, AstraZeneca, or Roche, who have utilized such assays to generate data submitted as part of the supporting information for the drug’s approval with regulatory bodies such as the FDA or the EMA.

Explore Examples of Approved Drugs Using Octet BLI^®