Affinity Characterization Service

For services that require the detection and monitoring of biomolecular interactions in real time, Kemp Proteins uses the ForteBio OctetRed96e system. The Octet system incorporates biosensors, reagents, and assay kits to enable us to perform high-throughput, real-time, label-free affinity and kinetic characterizations on a range of sample types including antibodies, proteins, and small molecules. Kemp’s highly experienced team and proven protocols deliver analyses based on solid scientific data, enabling our clients developing assays, diagnostic test kits and therapeutic products to make rapid decisions confidently.

The Octet RED96 is an instrument that can provide similar information as Biacore Surface Plasmon Resonance (SPR) using a detection different technology. The technology is called Bio-Layer Interferometry (BLI) Technology.

The OctetRed96e uses Bio-Layer Interferometry (BLI) technology for the evaluation of biomolecular interactions and is based on the optical interference pattern of white light reflected from a layer of immobilized protein and an internal reference layer. By monitoring interference patterns (binding events) in real time, our team calculates on rates (ka), off rates (kd), and affinity constants (KD) and obtains an association/dissociation curve by measuring the shift and plotting its size over time. Kemp’s ability to monitor binding specificity, association and dissociation rates, and concentration with outstanding precision and accuracy using a label-free, real-time, and high-throughput methodology provides a cost-effective alternative to Biacore. Let us help you with your affinity characterization project, request a quote now!

Antibody Characterization Services:

Let us help you with your Custom BLI / SPR Services project, request a quote now!

Types of Affinity Measurement Services:

Kemp provides the following antibody-related services using the OctetRed96e system:

Isotyping determination of antibody (IgG1, IgG2a etc.)
Quantification of IgG concentration in samples
Monoclonal antibody screening against label-free antigen in solution
Antibody pairing by “epitope binning” to identify and group different antibodies (to enable antibody pairing according epitope binding on the same antigen)
Full kinetic analysis/ ranking by on rates (ka), off rates (kd), and affinity constants (KD)

Common Ligand to Analyte Assay Formats:

Antigen and Antibody Fragment Characterization
Protein to Protein or Protein to Peptide Interactions
Protein to DNA or Protein to RNA interactions
Protein to Lipid Interactions
Protein to Small Molecule Interactions
Protein or Antibody Quantitation

Octet BLI vs Biacore SPR

	Octet	Biacore
Detection Technology	Bio-Layer Interferometry (BLI)	Surface Plasmon Resonance (SPR)
Signal Read Out	Wave Length Shift	Refractive Index
High Throughput	Yes – 96-well plate format allowing for multiple samples in a single run	No- 4 channel set-up. Limits the number of samples per run
Sample Recovery	Ability to fully recover samples for use in additional assays	Limited sample recovery as samples are consumed for analysis
Sample Analysis Time	As short as 24 minutes for 96 samples	Typically 3-4 hours for 3 samples
Costs per sample	Cost Effective	Relatively High

Case Study – Octet Kinetic Analysis

Experiment Summary Using AR2G biosensors

Procedures:

Binding experiments were performed on Octet RED96e at 30°C. Antibody1 was immobilized on AR2G biosensors, followed by binding of Ag (His Tagged Protein, 10kDa) at variable concentrations. Binding of antigen to the antibody was monitored in real time to obtain on (ka) and off (kd) rates. The equilibrium constant (KD) was calculated from the observed ka and kd.
Full kinetic analysis was performed using analyte concentrations from 1000nM to 4.1nM.

Case Study – Octet Epitope Binning

Procedures:

Binding experiments were performed on Octet RED96e at 30°C. The His-tagged protein was immobilized on the NiNTA biosensors. A label free Octet-based sequential antibody binding assay includes binding of the first mAb to the Ligand immobilized on the AR2G biosensor. A subsequent Ligand Binding step with a second mAb was performed to identify monoclonal antibodies that can simultaneously bind the Ligand. Epitope Bins are defined based on binding patterns. Antibody Pairs for sandwich immunoassays were identified for a sandwich immunoassay platform.

FAQ on Octet & Biacore Analysis:

What can I use Octet analysis for?

Characterization of high-affinity biologic interactions
Fc receptor binding assays
Kinetics characterization of large molecules
Protein: Small molecules and peptide kinetics & affinity characterization

If I have multiple antibodies, can you suggest pairs that will work in a sandwich ELISA?

Yes, we can use epitope binning to determine optimum pair(s) of antibodies for your assay.

Can you rank a number of antibodies using their kinetics?

Yes, we will perform full kinetics analysis to enable us to provide you with a list of antibodies ranked according to kinetics. (Specific rankings often depend on the projected use of the antibody.)

What types of screening can be performed for the projects?

Kemp Proteins utilizes automated high-throughput screening and analytical technologies to deliver monoclonal antibodies that consistently meet clients’ acceptance criteria. We perform Octet screening, high-throughput flow cytometry, Biacore analysis, direct ELISA, indirect/capture ELISA, and competition assays (blocking/non-blocking assays) at an additional cost.

What do you need to provide this analysis?

Sufficient quantity of test samples (antibody and antigen) based on the number of assays to be performed. Typically 150 ug for full kinetic analysis and 10-20 ug for binning.

Is there anything else required?

In addition to test samples, we will need the test sample’s molecular weight and its concentration as supplied.

How long does it take to obtain a report on my test samples?

Based on client-provided samples that have all the appropriate data, we can usually deliver a report the week following analysis.

What is the shipping address for Kemp Proteins?

Kemp Proteins, LLC

5119 Pegasus Court, Suite M-P

Frederick, MD 21704

Attention: Affinity Characterization Service

What do I receive?

Kemp Proteins will provide you with a report that contains the raw data (i.e., the graphic material provided by the Octet) and a summary of the findings for on rates (ka), off rates (kd), and affinity constants (KD), along with a ranking, if requested.

Why Kemp Proteins for Affinity Characterization Services?

Kemp Proteins experienced team of protein / antibody characterization scientists utilize modern characterization instrumentation and affinity protocols to deliver best in class reports and assays. Supported by a program management team and a Quality Management System that is ISO 9001 accredited, our experienced project managers use a process of pre-project consultation to ensure we understand your needs and expectations prior to starting any affinity project. We communicate with you throughout the entire process and deliver data packages and assays to help you succeed and advance your own programs. Our goal is to be the best Octet / Biacore Services company for our clients. We welcome any type of affinity project, we especially like the challenging ones!

Affinity Characterization (SPR/BLI) Service Quote Request

Once you complete and submit the form, one of our technical experts will will contact you within 1 business day to discuss your project. Thank you.

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