NuGel™ Polymer
Coated Silica Affinity Matrices For Affinity Purification of
Proteins.
Silica has been an industry standard
as an advantageous matrix suitable for high performance liquid
chromatography. Through a proprietary polymer coating, silica is
cross linked forming a reactive poly-epoxy functionality stable
across a wide pH range (pH 2 to 10). From this foundational
chemistry, all of the NuGel™
affinity products are derived.
- NuGel™
Poly-Epoxy has terminal epoxy as matrix reactive group and amino
group as the ligand reactive group with direct coupling of amino
groups.
- NuGel™
Poly-Amine has terminal amine as matrix reactive group and
carboxylic acid or carbohydrate group as the ligand reactive group
with carbodiiamide reaction or NaIO4 derived aldehyde
reaction.
- NuGel™
Poly-Aldehyde has terminal aldehyde as matrix reactive group and
amino group as the ligand reactive group with direct coupling of
amino groups.
- NuGel™
Poly-Hydroxy has terminal glycol as matrix reactive group and amino
as the ligand reactive group with carbodiiamidazole mediated
reaction.
- NuGel™
Poly -Carboxy has terminal carboxylic acid as matrix reactive group
and amino group as the ligand reactive group with carbodiiamide
mediated reaction.
- NuGel™
Poly -NHS has terminal N-Hydroxy succinimide as matrix reactive
group and amino group as the ligand reactive group with direct
coupling of amino groups.
NuGel™
polymer coated silica affinity matrices has applications for enzyme
immobilization, separation and purification of biopolymers,
immobilization of proteins & ligands – monoclonal antibodies,
hormones, peptides, haptens, drugs, etc.
Applications of NuGel™ Affinity Matrices
Ehrlich, G. K., Michel, H., Chokshi, H. P. and Malick, A. W. Affinity purification and characterization of an anti-PEG IgM. Journal of Molecular Recognition, 22: 99–103 (2009).
Anti-PEG IgM Purification : Scientists used PEG-derivatized NuGel™ to bind anti-PEG IgM for purifying anti-PEG IgM. Subsequent analysis of anti-PEG IgM and fragments occurred using PEG functionalized proteins/peptides by surface plasmon resonance, western blotting and ELISA. Such studies create probes for assays used in functionalized PEG molecules.
Membrane-based receptor affinity chromatography. Journal of Chromatography A Volume 597, Issues 1-2, 24 April 1992, Pages 155-166 9th International Symposium on Affinity Chromatography and Biological Recognition
Receptor Affinity Chromatography: In Purification and Analysis of Recombinant Proteins authors Seetharam and Sharma cite NuGel™ affinity support immobilization which allows for efficient receptor coupling and recovery of binding capacities. Authors recommend NuGel™ for industrial scale operations because of mechanical rigidity and flow rates which provide a durable bed support resulting in four fold flux than compared to agarose supports. Authors present the use of receptor affinity chromatography of recombinant interleukin 2 for the purification of recombinant proteins.
Purification of immunoglobulins using affinity chromatography and peptide ligands. Ruben Carbonell et al US Patent 2006/0153834 A1
Affinity Purification Of Human Immunoglobulins Using Immobilized Peptide Ligands on NuGel™ Poly-Amine Support: NuGel™ Poly-Amine is used for the purification of human immunoglobulins (Igs), a class of plasma proteins produced by the immune system.
George K. Ehrlich, Pascal Bailon, Wolfgang Berthold. Phage Display Technology - Identification of Peptides as Model Ligands for Affinity Chromatography Affinity Chromatography Methods in Molecular Biology, 2000, Volume 147, 209-220
Screening of Phage Peptide Libraries:
In the procedure for screening of phage peptide libraries against monoclonal antibodies, scientists used NuGel™ for coupling peptide mimotope to the affinity matrix for purifying antibodies from human serum. Producing peptides on bacteriophages requires selection for ligand-binding molecules ie selection against target molecules and tailoring ligands to numerous applications. Authors Ehrlich et al describe the selection of peptide ligands for affinity chromatography (especially peptides that mimic antigen epitopes).
References:
Patents
Monoclonal antibodies directed to the cytotoxic lymphocyte maturation factor European Patent EP0790255
Purification of immunoglobulins using affinity chromatography and peptide US 2006/0153834 A1
Affinity
Dermot Walls, Robert McGrath and Sinéad T.Loughran A Digest of Protein Purification. Methods Molecular Biology. Volume 681: 3-23 (2011)
Ehrlich, G. K., Michel, H., Chokshi, H. P. and Malick, A. W. Affinity purification and characterization of an anti-PEG IgM. Journal of Molecular Recognition, 22: 99–103 (2009).
Development of hepatitis B virus capsids into a whole-chain protein antigen display platform: New particulate Lyme disease vaccines. International Journal of Medical Microbiology Volume 298, Issues 1-2, 3 January (2008), Pages 135-142
A sensitive and high-throughput assay to detect low-abundance proteins in serum Hongtao Zhang, Xin Cheng, Mark Richter & Mark I Greene. Nature Medicine 12, 473 - 477 (2006)