Concanavalin A immobilized magnetic poly(glycidyl methacrylate) beads for antibody purification

Abstract

Concanavalin A (Con A) immobilized magnetic poly(glycidyl methacrylate) (mPGMA) beads in monosize and spherical for (1.62 μm in diameter) were used for the purification of human immunoglobulin G (IgG) from human plasma. Con A was immobilized by covalent binding onto the mPGMA beads. The maximum IgG adsorption on the mPGMA‐Con A beads was observed at pH 6.0. The nonspecific IgG adsorption onto the plain mPGMA beads was very low (0.22 mg/g). Scatchard analysis of the adsorption isotherm for IgG on mPGMA‐Con A beads showed an affinity constant (Ka) of 1.39 × 105 M−1 and a theoretical maximum adsorption capacity of 109.1 mg/g. An apparent IgG adsorption capacity of 66.2 mg/g was observed under the experimental conditions. IgG adsorption capacity from human plasma was observed as 48.0 mg/g. The adsorption of human serum albumin from plasma was 2.0 mg/g. The total protein adsorption was determined to be 50.0 mg/g. IgG molecules could be repeatedly adsorbed and eluted with the mPGMA‐Con A beads without noticeable loss in the IgG adsorption capacity. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Polyethyleneimine-modified graphene oxide nanocomposites for effective protein functionalization

“…After selective enrichment by GO@BPEI@Con A, unwanted peaks of Con A (dyadic monomer peak of Con A at 12.5 kDa) and Myo can not be detected in the spectra, indicating the tightly covalent linkage between Con A and GO@BPEI and the complete removal of non-glycoproteins. [45][46][47] In addition, the non-glycoproteins were barely absorbed on GO@BPEI@Con A, confirming the high activity and specificity of the immobilized Con A. Interestingly, the selective adsorption capacity of GO@BPEI@Con A for different glycoproteins varied in their varieties. For comparison, commercial agarose-Con A was also utilized to capture RNase B from the protein mixture of RNase B and Myo (1:500, m/m), followed by MALDI-TOF MS analysis.…”

Recent Advances in Lectin-Based Affinity Sorbents for Protein Glycosylation Studies

“…Home-Made Lectin-Based Sorbents for Dispersive Solid-Phase Extraction Home-made lectin-based sorbents based on polymeric particles (Helmholz et al, 2003;Yavuz et al, 2004;Rosenfeld et al, 2005;Aniulyte et al, 2006;Niemeyer, 2011, 2011;Hajizadeh et al, 2012;Demir et al, 2018) were applied also in dSPE, as summarized in Table 5. Additionally, dSPE processes can also be performed with lectin grafted to magnetic particles (Ferreira et al, 2011;Akkaya et al, 2012;Qin et al, 2014;Idil et al, 2015;Miura et al, 2018;Waniwan et al, 2018). This approach can simplify the separation process between the bound and the unbound fraction, as it necessitates the use of a magnet to separate both fractions rather than by a tedious centrifugation step.…”

Lectin Affinity Based Recognition Nanomaterial for Glucose

“…In a study, it was determined that the amino acids Asp 81, Gly 99, Asn 125, Ala 30, Phe 123 and Glu 31 were involved in the binding region of Con A. It has been found that these amino acids form a network of hydrogen atoms with the oxygen atoms in the hydroxyl groups bound to the 3rd, 4th, 5th, and 6th carbon groups of mannose and glucose sugars [15,16].…”

“…Concanavalin A (Con A) is a typical lectin protein, a tetramer at physiological pH (7.4) comprised of four identical monomeric subunits (MW ∼ 25 kDa). Concanavalin A (Con A) is also a metalloprotein containing Mn 2+ and Ca 2+ [16]. Concanavalin A (Con A) has been extensively used in the isolation, fractioning and structural characterization of glycoproteins and other important glycoconjugates carrier glucose and/or mannose residues [3][4][5].…”