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BLI Affinity Determination Services

KMD Bioscience provides accurate and rapid affinity determination service for customers based on ForteBio Octet system. BLI (Biolayer Interferometry) is a label-free and fluidics-free, real-time measurement and monitoring system based on light intensity interference. ForteBio Octet platform is shown in Figure 1.

Traditional molecular interaction (Co-Immunoprecipitation, Pull Down, EMSA, ELISA, etc.) not only can get qualitative results, but BLI can get quantitative results. Due to the high flexibility and sensitivity of multiple sensors, researchers can conduct a wide range of studies, including antigen and antibody affinity measurements, protein and peptide affinity measurements, protein and small-molecule drug affinity measurements, and so on.

KMD Bioscience's technical support will design a detailed experimental plan for you according to your needs. We will provide accurate and effective consulting service based on BLI technology before starting a new project. With our advanced ForteBio Octet system, KMD Bioscience can deliver analysis data with high accuracy and reasonable price.

 

Figure 1 ForteBio Octet Platform

 

Biolayer Interferometry Technology(BLI):

 

The principle of BLI is that a beam of visible light passes through an optical fiber, and two beams of reflection spectrum are formed at the two interfaces of the optical film layer at the sensor end, which superposes to form a beam of interference spectrum. Molecular binding leads to changes in the thickness of the film layer, which is reflected by the displacement value of the interference spectrum. The schematic diagram is shown in Figure 2.

 

Figure 2 BLI Principle

 

Application:

 

KMD Bioscience provides molecular interaction service by BLI.

BLI technology has a wide range of applications, including small molecule (<100 Da), protein, nucleic acids, lipids, bacteria, viruses and whole cells.

Many studies have applied BLI technology to rapidly realize affinity determination, protein quantitative analysis and kinetic analysis. For example, ZHOU L J et al. reported the crystal structure of the heptameric Lsm1-7 complex and preliminary characterization of its RNA-binding properties by BLI.LIU C X et al. determined the binding rate constants of peroxidase I and peroxidase II to biotinylated adenosine at different time points by BLI, they show that adenanthin directly targets the conserved resolving cysteines of Prx I and Prx II and inhibits their peroxidase activities. KAHSAI A W et al. used BLI technology to screen RNA aptamers against G protein-coupled receptors. Significantly expanding the diversity of ligands that can be used to study the regulation of structure and function of G protein-coupled receptors. WANG W J et al. carried the kinetic analysis of affinity of WT or mutant SUN domains and KASH domain by Octet Red 96. Results showed that BLI technology can not only detect affinity, but also reflect the binding mode of the complex. DZIMIANSKI J V et al. described a novel application of biolayer interferometry for the rapid detection of antigen-specific antibody levels in plasma samples and demonstrated its utility for quantification of SARS-CoV-2 antibodies. Using biochemical approaches, WEI Y et al. showed that MYC amino acids 115–124 also interact with TBP independently of TAF1TAND1. WANG Z J et al. performed BLI experiments in which a preformed antibody–RBD complex was exposed to a second monoclonal antibody targeting one of three classes of structurally defined epitopes. To better quantify peptide binding to BAZ1A-BD, OPPIKOFER M et al. used biolayer interferometry to measure its interaction with a tetraacetylated H4 1–19 peptide.

BLI technology can also be used for the study of protein function. KAWAHARADA Y et al. showed that the extracellular domain of EPR3 can specifically recognize monomeric EPS by BLI technology. With the help of BLI technology, LANDGRAF K E et al. used SA sensors to study multi-molecular synergy, which has broad significance for the development of selective allosteric activators of serine proteases and pseudoproteases. 

KMD Bioscience uses high-sensitivity instruments and provides precise experimental plans, which can effectively help customers carry out research on small molecule, lipid and nucleic acid and obtain quantitative data.

--Measurement throughput: 96 channels

--Molecular measurement limit:150 Da

--KD Range:1 mM~10pM

--Sample Range: Serum, Cell lysate, Supernatant, Fermentation broth, Tissue homogenate and other crude samples, Purified samples.

--Kon(Association Rate):102~107M-1S-1

--Koff(Dissociation Rate):10-6~10-1S-1

 

 Determination Type:

 

BLI kinetic assays are commonly used in the presence or absence of specific binding between samples, binding affinity, on/off rates, screening and sorting, competition experiments, epitope pairing and other kinetic assays (Figure 3); quantity of antibody or recombinant, measurement of antibiotics or toxins, ELISA quantitative alternative experiment and other concentration measurement (Figure 4). Each assay type generates unique information that aids in the analysis of biomolecules.

Figure 3 BLI Kinetic Determination Curve

Figure 4 BLI Concentration Determination Curve

 

Service Range:

 

KMD Bioscience provides label-free molecule interaction service based on BLI technology: Protein-Protein affinity detection; Antigen-Antibody affinity detection; Protein-Small molecule drugs affinity detection; Protein-DNA affinity detection; Antibody isotyping analysis; Antibody pairing; hybridoma screening, etc. As well as many other compounds and high-throughput affinity sequencing services according to customer requirements.

We have more than ten kinds of sensors to meet various needs of different samples. According to different experimental samples and purposes, we will choose matched sensors for experiments.

 

Sensor Type

Sensor Name

 

 

Common Type

Streptavidin (SA)

Super Streptavidin (SSA)

High Precision Streptavidin (SAX)

* Amine Reactive 2nd Generation (AR2G)

* Aminopropylsilane (APS)

 

Capture Type

* Anti-Penta-HIS(HIS)

Ni-NTA(HIS)

* Anti-GST

 

 

 

 

Antibody Type

* Anti-Human IgG Fc (AHQ)

Anti-Human IgG Fc (AHC)

Anti-Murine IgG Fv (AMQ)

* Anti-Murine IgG Fc (AMC)

* Anti Human IgG Fab (FAB2G)

* Protein A

* Protein G

* Protein L

 

Sample Requirements:

 

Customer provides

Requirements

 

 

Customer can provide antibody/protein/peptide/small molecular compound etc.

* Buffer: PBS, HEPPS etc. Try not to contain glycerol, imidazole, trehalose or other salts; try to avoid reagents with amino groups such as Tris as buffer.

* Protein>2 mg, Concentration>0.5 mg/ml

* Antibody>200μg, Concentration >0.5 mg/ml

* Peptide>200 μg,Concentration >0.5 mg/ml

* Compound small molecule>1 mg, Concentration >1 mg/ml

 

Service Highlights:

 

--Technical support provides professional service, meet customer's requirements.

--High accuracy by professional operation and strict quality control.

--High efficiency, label-free, real-time, high-throughput affinity characterizations by using advanced Octet system.

--Method design is more convenient, simply, flexible and easy to operate.

--Higher measurement sensitivity and lower sample consumption.

--Various sensors.

--Strong acceptance: could apply to crude samples, such as cell supernatant, culture medium.

--Strong anti-interference ability: temperature, buffer refractive Index don't affect the experiment.

--Wide range: protein, nucleic acid, peptide, nanoparticles etc.

--One-stop downstream services, providing integration from affinity measurement service to molecule interaction.

--Rich experience in affinity and kinetic detection.

 

How to Order?

 

If you have any questions regarding our services or products, please feel free to contact us by E-mail: info@kmdbioscience.com or Tel: +86-400-621-6806, We will reply as soon as possible.

 

Reference

[1] ZHOU L J, ZHOU Y L, HANG J, et al. Crystal structure and biochemical analysis of the heptameric Lsm1-7 complex [J].

[2] LIU C X, YIN Q Q, ZHOU H C, et al. Adenanthin targets peroxiredoxin I and II to induce differentiation of leukemic cells [J].

[3] KAHSAI A W, WISLER J W, LEE J, et al. Conformationally selective RNA aptamers allosterically modulate the beta(2)-adrenoceptor [J].

[4] WANG W J, SHI Z B, JIAO S, et al. Structural insights into SUN-KASH complexes across the nuclear envelope [J].

[5] DZIMIANSKI J V, LORIG-ROACH N, O'ROURKE S M, et al. Rapid and sensitive detection of SARS-CoV-2 antibodies by biolayer interferometry [J].

[6] WEI Y, RESETCA D, LI Z, et al. Multiple direct interactions of TBP with the MYC oncoprotein [J].

[7] WANG Z J, MUECKSCH F, SCHAEFER-BABAJEW D, et al. Naturally enhanced neutralizing breadth against SARS-CoV-2 one year after infection [J].

[8] OPPIKOFER M, SAGOLLA M, HALEY B, et al. Non-canonical reader modules of BAZ1A promote recovery from DNA damage [J].

[9] KAWAHARADA Y, KELLY S, NIELSEN M W, et al. Receptor-mediated exopolysaccharide perception controls bacterial infection [J].

[10] LANDGRAF K E, SANTELL L, BILLECI K L, et al. Allosteric Peptide Activators of Pro-Hepatocyte Growth Factor Stimulate Met Signaling [J].