Protein Purification

 

Gentle protein purification using patented MAB-SensBeads®

Gentle protein purification when using MAB-SensBeads®, novel crosslinking chemistry and neutral high salt conditions:

  • very pure protein eluates
  • high protein recovery
  • elution under neutral conditions reducing denaturation of protein and antibody
  • no streptavidin-coated beads and biotinylated antibodies necessary
  • no protein A/G-beads and crosslinking necessary 
  • Normally, magnetic particles are comprised of organic polymers.

To minimize unspecific protein binding, particle surfaces must be coated with proteins. These protein coatings are often not stable under harsh elution conditions like acids, organic solvents or high salt conditions, which are required for during immunopurification of proteins.

Different strategies are possible:

 

1.) Drawback when using streptavidin-particles and biotinylated antibodies:


The antibody of interest must be biotinylated and purified after biotinylation, resulting in antibody loss.
Streptavidin coatings and streptavidin by itself is not stable under harsh elution conditions.

2.) Drawback when using Protein A/G:

The antibody of interest must be crosslinked to Protein A/G resulting
in a possible lower binding capacity of the antibody.
Suitable crosslinkers are often not stable under harsh elution conditions
(loss of antibody).

3.) MAB-SensBeads®

 The novel MAB-SensBeads® show, as compared to other silica particles,
very low unspecific protein binding and higher antibody binding capacity.

4.) Novel elution process

 A novel elution process under neutral high salt conditions reduces unspecific proteins in the eluates (Eluent B) as compared to normally used acid conditions.

Comparison of MAB-SensBeads® with other particles and methods:



 


 

  • 1 Protein marker.
  • 2 Streptavidin-coated particles (Distributor 1) + monobiotinylated PSA antibody.
  • 3 Protein G-coated particles (Distributor 2) + PSA antibody and
    crosslinker 1. 
  • 4 MAB-SensBeads® + PSA antibody, coupled with crosslinker 2.
  • 5 MAB-SensBeads® + PSA antibody, coupled with crosslinker 1.