Protein retention in dextran-grafted cation exchange chromatography:The influence of pHs,counterions and polymer structure

Polymer-grafted ion exchange adsorbents were of great interest for the development of high-performance protein chromatography in biopharmaceutical and related fields.In this work,protein retention was systematically investigated in ion exchange chromatography packed respectively with dextran-grafted cation exchange adsorbents containing sulphopropyl(SP)ligand,SP Sepharose XL and Capto S,and non-grafted cation exchange adsorbent,SP Sepharose FF,using five proteins.With an increase of buffer p Hs,retention factors of proteins decreased among all the adsorbents,demonstrating the dominant role of electrostatic interaction for protein binding on cation exchange adsorbents.The evidences further revealed that the scattered positive charges on the surface of protein molecules,rather than net charge of protein molecule,determined protein retention on cation exchange adsorbent.Likely,counterions including NH4^+,K^+,Na^+and Mg^2+exhibited distinct influence on protein retention.It was well ascribed to solvent-mediated indirect ion-macromolecule interactions and direct ion-macromolecule interactions.Compared with SP Sepharose FF,polymer structure in dextran-grafted cation exchange adsorbents ultimately brought about different ligand distributions and smaller pore sizes,thereby regulating protein retention in cation exchange chromatography.By comparing the retention of myoglobin andβ-lactoglobulin B in SP Sepharose XL and Capto S,we reasonably speculated that the enhancement of nonelectrostatic interaction caused by reducing the space arm length was a major reason for an increasing retention factor of myoglobin in Capto S.The results in this research help us understand adsorption mechanism of protein in polymer-grafted adsorbents and give scientific guidance for the development of chromatographic materials.

Effects of Dietary Protein Level on Growth and Utilization of Protein and Energy by Juvenile Mangrove Red Snapper(Lutjanus argentimaculatus)

A feeding trial was conducted in a recirculating water system to investigatethe effects of dietary protein levels on growth, feed utilization, hepatosomatic index and liverlipid deposition of juvenile red snapper, Lutjanus argentimaculatus (average initial wet weight 8.0± 0.39 g and total length 3.14 ± 0.3 cm). In the experiment, six fishmeal-based diets wereformulated to contain various protein levels (20% to 45% in 5% increments), with dietary energyranging from 2210.7kJ100g to 2250.2 kJ 100g dry matter. The protein to energy ratios of diets rangedfrom 8.58 mg protein kJ^(-1) to 20.03 mg protein kJ^(-1). Diets were fed for 90 d to triplicategroups of fish stocked in 0.128 m^3 seawater tanks, 25 individuals each. The daily ration of 2% wetbody weight was offered to the fish thrice a day. The fish at the end of the study had more thanten-fold (77.0 g) increase in weight compared to the initial (8.0 g). Fish fed diets of 40% and 45%protein produced significantly (P【0.05) higher weight gain of 77.2 g and 76.5 g, and specific growthrate (SGR) of 2.65% and 2.62% than those of 67.0 g and 68.3 g, and 2.49% and 2.51% of the otherdiets. The broken-line regression of SGR against dietary protein level yielded an optimum dietaryprotein requirement of 42.6% (Y = - 1.6295 + 0.1114 X^2, P【0.05). Survival remained 100% amonggroups. Feed conversion ratio decreased from 0.45 for fish fed 20% dietary protein to 0.35 for fishfed 45% dietary protein. Nitrogen intake increased with an increase in dietary protein, which inturn resulted in an increase in nitrogen gain of fish whole body. Fish fed 40% and 45% protein dietsshowed higher (P【0.05) nitrogen gain (0.27g and 0.26g) than those (0.23g and 025g) fed all otherdiets. Gross energy intake (GEI) in fish fed 45% protein was lower (600.67kJ) than that (607.97 kJ)of 40% protein diet, though the differences were not statistically significant (P】0.05); GEI rangingfrom 677.31 kJ to 663.20 kJ at remaining four diets (20% to 35% protein) did not appear to differsignificantly (P】0.05). The highest energy gain of 518.33 kJ was obtained with fish fed 40% protein,resulting in the highest energy retention efficiency of 85.26%. The hepatosomatic index of fish feddiets of 20%, 25%, 30% and 35% protein were significantly (P【0.05) higher (2.09% to 2.57%) thanthose (1.44% and 1.41%) of fish fed diets containing 40% and 45% protein. Liver lipid contentsdecreased from 8.72% to 7.0% in fish fed dietary protein of 20% to 45% in 5% increments. Resultssuggest that the diet containing 40% to 42.6% protein with a P/E ratio of 17.6 mg protein kJ^(-1) isrequired for good growth of L. argentimaculatus weighing between 8.0 g and 85.2 g under the cultureconditions of the present study.