Maxwell® Antibody Capture System

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Rapid, High-Throughput Automated Antibody Purification

  • Increased binding capacity, excellent analyte recovery and minimal nonspecific binding
  • Wide, linear dynamic range
  • All required bulk reagents included for enrichment of biotherapeutic proteins from animal matrices
  • Fully compatible with a wide range of automated liquid handling platforms

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Maxwell® Antibody Capture System

Automated Purification of Biotherapeutic Proteins

Antibody purification from a large number of complex biological matrices (e.g., serum, plasma, cell media, etc.) is often the first step in the characterization of therapeutic antibodies using methods such as liquid chromatography/tandem mass spectrometry (LC-MS/MS). We have developed a solution for rapid, simple, high-throughput (HT) automated purification of antibodies with our Maxwell® HT Ab Capture Kit (for KingFisher® and Automated Liquid Handling Platforms).

Hybrid Immunoaffinity (IA) LC-MS/MS Assay Workflow

Hybrid Immunoaffinity (IA) LC-MS/MS Assay Workflow

Conceptual overview of the Hybrid IA LC-MS/MS workflow (enrichment step). Following enrichment, a proteolysis step (usually with trypsin) is required prior to LC-MS/MS analysis.


Therapeutic Antibody Purification

High-Throughput (HT) Purification

The Maxwell® HT Ab Capture Kit, containing High Capacity Magne® Streptavidin Beads and all critical reagents, is now available.

  • Method development IgG1 and SIL IS IgG1 included
  • Flexibility for user-provided Antibody and Standard
  • Compatible with various bead processors and high-throughput (HT) liquid handlers
  • Detailed protocol and turnkey methods available upon request

For customers using Thermo Fisher KingFisher® magnetic bead processors or liquid handlers in their laboratories, the Maxwell® HT Ab Capture Kit can be used immediately to isolate biotherapeutic proteins prior to proteolysis.

Overview of the therapeutic antibody purification workflow

Overview of the simple therapeutic antibody purification workflow. Prepare a 96-well plate with samples (calibration, QC, unknowns, etc.) and add beads. Allow for 2 hours of binding. Wash and elution steps are carried out with either a magnetic bead processor or liquid handler.

Why Use High Capacity Magne® Streptavidin Beads?

Improved Binding Capacity

Recovery of Humira from various sources using high-capacity Magne Beads.

Recovery of Humira spiked into various matrices. Following binding on the Eppendorf MixMate® instrument, samples were processed using the KingFisher® instrument (Thermo Fisher). Panel A. Recoveries of Humira using the High Capacity Magne® Streptavidin beads were 80% for the non-CA (cross-adsorbed), 88% for rodent and 99.7% Cynomolgus (Cyno). Panel B. Compared with Competitor D, 90% improvement in binding capacity was observed.

Improved Assay Window

Calibration curves of immunoaffinity magnetic capture beads

Calibration curves of immunoaffinity magnetic capture beads. Calibration curves were generated from samples processed using High Capacity Magne® Streptavidin Beads (Panel A) or beads from Competitor D (Panel B). Panel A. Linearity was observed without internal standard normalization and low variance of Internal standard (IS) peak area (Panel C, 9.3%). Panel B. Competitor D beads fail to show linearity across the calibration range (50μg/ml to 25ng/ml) and high variance of IS peak area (Panel D, 26%). The blue arrow refers to a strong deviation in the IS area intensity for the high-concentration samples. Peaks areas across the assay should be less than 15%.


Method Validation

The Hybrid Immunoaffinity LC-MS/MS Assay was evaluated on the KingFisher® System. The assay shows good linearity, precision and accuracy.

Hybrid Immunoaffinity LC-MS/MS assay linearity, precision and accuracy on the Kingfisher System.

Assay linearity, precision and accuracy. Good precision and accuracy (well within GLP guidelines) was observed on the KingFisher® System.

What’s in the box?

Maxwell® HT Ab Capture Kit

  • 1X Binding Buffer, 400ml
  • 1X Wash Buffer, 700ml
  • 1X Elution Buffer, 50ml
  • Bead Dilution Buffer, 400ml
  • High Capacity Magne® Streptavidin Beads, 11ml
  • Method IgG, 4mg
  • SIL IS IgG, 0.4mg

Researchers will need to supply biotinylated capture antibodies due to the wide range of varying analytical targets. A non-cross-adsorbed capture antibody is available from Promega (Cat.# V7830).

Sufficient reagents are included for four plates.

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