BIOMAT Database Logo BIOMAT Database Logo

High throughput purification of monoclonal recombinant antibodies using a Protein-A coated membrane plate system. 2023

Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
New Mexico Consortium, 4200 W. Jemez Rd, Suite 301, Los Alamos, NM 87544, USA.

The therapeutic use of monoclonal antibodies (mAbs) ranges from cancer treatment to immune-mediated conditions, covering infectious and cardiovascular disorders, among others. The development of improved methods for therapeutic antibody discovery has accelerated the identification of numerous mAbs: a discovery campaign can be deeply mined, resulting in hundreds, even thousands, of potential antibody leads for a given target of interest. High throughput mAb expression and purification methods are required for the rapid validation of those leads. In this work, we describe the implementation of a Protein-A coated membrane plate system, the Purexa™ AHT membrane plate, for robust preparative purification of hundreds of recombinant mAbs, without the need for automation. The high efficiency (>80%) recovery generated sufficient mAb for downstream screening analyses such as ELISA and surface plasmon resonance (SPR). This new system allows the functional validation of hundreds of lead antibodies from discovery campaigns in a timely manner regardless of operational size.

UI MeSH Term Description Entries
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D013205 Staphylococcal Protein A A protein present in the cell wall of most Staphylococcus aureus strains. The protein selectively binds to the Fc region of human normal and myeloma-derived IMMUNOGLOBULIN G. It elicits antibody activity and may cause hypersensitivity reactions due to histamine release; has also been used as cell surface antigen marker and in the clinical assessment of B lymphocyte function. Protein A,Protein A, Staphylococcal
D020349 Surface Plasmon Resonance A biosensing technique in which biomolecules capable of binding to specific analytes or ligands are first immobilized on one side of a metallic film. Light is then focused on the opposite side of the film to excite the surface plasmons, that is, the oscillations of free electrons propagating along the film's surface. The refractive index of light reflecting off this surface is measured. When the immobilized biomolecules are bound by their ligands, an alteration in surface plasmons on the opposite side of the film is created which is directly proportional to the change in bound, or adsorbed, mass. Binding is measured by changes in the refractive index. The technique is used to study biomolecular interactions, such as antigen-antibody binding. Plasmon Resonance, Surface,Plasmon Resonances, Surface,Resonance, Surface Plasmon,Resonances, Surface Plasmon,Surface Plasmon Resonances

Related Publications

Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
Development of a high throughput protein a well-plate purification method for monoclonal antibodies.
January 2009, Biotechnology progress,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
Engineering Staphylococcal Protein A for high-throughput affinity purification of monoclonal antibodies.
November 2020, Biotechnology advances,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter.
May 2021, Journal of visualized experiments : JoVE,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
A high-throughput purification of monoclonal antibodies from glycoengineered Pichia pastoris.
November 2010, Protein expression and purification,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
A platform for high-throughput molecular characterization of recombinant monoclonal antibodies.
November 2005, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
Purification of monoclonal antibodies using protein a/g.
January 2000, Methods in molecular medicine,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
High-throughput process development for recombinant protein purification.
March 2006, Biotechnology and bioengineering,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
Studying host cell protein interactions with monoclonal antibodies using high throughput protein A chromatography.
October 2012, Biotechnology journal,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
A multi-column plate adapter provides an economical and versatile high-throughput protein purification system.
December 2018, Protein expression and purification,
Camila Leal-Lopes, and Sara D'Angelo, and M Frank Erasmus, and Andre A R Teixeira, and Graham Temples, and Jinxiang Zhou, and Andrew R M Bradbury, and Fortunato Ferrara
A centrifugation-free high-throughput protein purification system using in-line microfluidization.
October 2011, Protein expression and purification,
Copied contents to your clipboard!