Poly(Hydroxyethyl Methacrylate) Immunoaffinity Cryogel Column for the Purification of Human Immunoglobulin M

Abstract

Human immunoglobulin M (hIgM) antibodies are considered as hopeful tools for diseases therapy. Therefore, chromatography approaches are used to purify hIgM with a single step. In this study, we prepared a poly(hydroxyethyl methacrylate) based immunoaffinity p(HEMA-I) cryogel column by using cyanamide to immobilize antihuman immunoglobulin on the p(HEMA) cryogel for purification of hIgM in aqueous solution and artificial human plasma. The characterization of the p(HEMA) cryogel column was performed by using a scanning electron microscope (SEM), micro-computerized tomography (µ-CT), Fourier transform infrared spectroscopy (FTIR), swelling degree and macro-porosity. Further, the optimizations of various parameters were performed such as, pH, ionic strength, temperature and concentration of hIgM in aqueous solutions. In addition, the Langmuir adsorption model was supported by experimental results. Maximum adsorbed amount of hIgM corresponded to 11.1 mg/g at pH 5.75 [morpholino ethanesulfonic acid (MES buffer)]. Our results indicated that the p(HEMA-I) cryogel column can be reused at least 10 times without significant loss in adsorption capacity. As a natural source, artificial human plasma was selected for hIgM adsorption and the purity of hIgM was evaluated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Preparation of Boronic Acid-Functionalized Cryogels Using Modular and Clickable Building Blocks for Bacterial Separation

“…Three replications were made in each case. The swelling degree of the cryogels was determined using the equation 40 , 41 where W 1 and W 2 are the weight of the swollen and the dry cryogel, respectively.…”

“…In the next step, the swollen cryogel was squeezed to remove the water, and the weight of the squeezed cryogel ( W 3 ) was measured again. The macroporosity (%) of the cryogel was determined based on water vapor adsorption and calculated using the equation 41 where ρ H 2 O and ρ gel are the density of water at 25 °C and the density of swollen cryogel, respectively.…”

Surface Plasmon Resonance-Based Immunosensor for Igm Detection with Gold Nanoparticles

“…IgM is the largest (950 kDa) and the first produced antibody in human serum to combat or stop the infections with the higher avidity of the antigens because of having 10 free antigen-binding sites [ 1 , 2 , 3 , 4 ]. The monitoring of IgM level in biological fluids is helpful to diagnose bacterial or viral infections from Dengue virus to some of certain diseases; including, acute chronic hepatitis, rheumatoid arthritis; furthermore, IgM deficiency is an immune disorder related to serious infections [ 2 , 5 , 6 , 7 , 8 ].…”

Molecularly imprinted cryogel cartridges for the selective recognition of tyrosine

“…Cryogel can be used in the purification of molecules. [5][6][7] Cryogel is an alternative to imprinting supports because it possesses many advantages such as large pores, low-pressure drop, short diffusion path, and very short residence time for both adsorption and elution. Therefore, this unique material can be used for the selective purification of amino acids.…”

“…Cryogel is gel matrices formed and has a great deal of interest in biomedical, biotechnology, and pharmacy. Cryogel can be used in the purification of molecules 5‐7 . Cryogel is an alternative to imprinting supports because it possesses many advantages such as large pores, low‐pressure drop, short diffusion path, and very short residence time for both adsorption and elution.…”