MBP purification by affinity chromatography with nanoCLAMP malE-A1(Resin), batch mode from E. coli lysate


Overview
Abstract
MBP purification by affinity chromatography with nanoCLAMP malE-A1(Resin) in batch mode from E. coli lysate. E.coli were lysed with BPER (Thermo), cleared by centrifugation, and diluted with PBS to 1.2 mg total protein/ml. The lysate was spiked with recombinant antigen to 0.1 mg/ml in 1.5 ml and incubated with 10 µl (packed vol) malE-A1(Resin). Beads were washed with PBS and eluted with polyol elution buffer at near neutral pH.
Type
  • Affinity Chromatography
Specimen
Target
Scores
Outcome
Successful
Signal
Acceptable
Cross Reactivity
Acceptable
Background
Acceptable
Dynamic Range
Acceptable
Results
Summary
MBP was successfully purified from an E.coli whole cell lysate to apparent homogeneity in one chromatography step.
Figure

Image 

MBP purification by nanoCLAMP malE-A1 affinity chromatography

Legend 

SDS-PAGE analysis of affinity purification of MBP from E.coli BL21DE3 whole cell lysates using malE-A1(Resin) affinity resin. E.coli were lysed with BPER (Thermo), cleared by centrifugation, and diluted with PBS to 1.2 mg total protein/ml. The lysate was spiked with recombinant antigen to 0.1 mg/ml in 1.5 ml and incubated with 10 µl (packed vol) malE-A1(Resin). Beads were washed with PBS and eluted with polyol elution buffer at near neutral pH. Lysates and eluants were analyzed on 12% SDS-PAGE in reducing SDS sample buffer.

Lane M: 10 µl See Blue Plus pre-stained protein marker (Thermo).

Lane 1: MBP-spiked lysate containing 21 µg of total protein.

Lane 2: 16 µl of polyol eluant from the affinity resin, from 121 µl total elution.