Introducing mabselect prisma, the nextgeneration protein a chromatography resin offering significantly enhanced alkaline stability and binding capacity for improved bioburden control and process economy in mab processing. The breakthrough capacity q b, 10% and additional stability indicating parameters were measured at 0, 6, 12, 24, and 36 months. Mabselect sure superior resistance and mabselect sure superior resistance lx with novel, alkalitolerant recombinant protein a ligand that provides greater stability and allows use of 0. The capacity of conventional protein a resin mabselect is also included in the figure for comparison. Relativedynamicbindingcapacity,qb10%,formabselectprismaafter cipwith0. Here, a fixed column size of 450200 mm, a mab titer of 4 gl, and a 20% safety factor in loading were used. Mabselect sure lx has been further developed from mabselect sure to. Data file 11003490 ac affinity chromatography hitrap mabselect sure hitrap mabselect hitrap mabselect xtra fig 1. Dynamic binding capacity study on mabselect sure lx for. Application note 28907892 ad processscale antibody. Here, we provide an overview of a novel system for mab capture from a users perspective.
Typical pressureflow profile for mabselect sure in a packed bed bed height 20 cm in a bpg 300 column i. Toyopearl afrprotein a hc650f toyopearl afrprotein a650f mabselect sure lx column size. The latest member of ge healthcares mabselect family is called mabselect sure lx. Plate number, assymetry and pressure vs flow were determined. Mabselect sure lx is a protein a affinity resin with very high dynamic binding capacity at extended residence times. Mabselect protein a resins ge healthcare life sciences. Mabselect prisma affinity columns and resins for antibody. The protocol includes a cip procedure to minimize crosscontamination from different antibodies between purification runs, as well as for general column cleaning after a purification run. Even more marked results were obtained with a papain digest of a commercial v h 3 antibody, herceptin roche products, which also showed coelution of fab and fc on the kitprovided proteina but separation on mabselect sure fig. Mabselect sure lx and mabselect sure were packed in tricorn 5100 columns bed height 10 cm, column volume 1. Mabselect sure lx is developed for hightiter mab capture at process scale. Mabselect prisma in comparison with mabselect sure and mabselect sure lx. As a result, mabselect sure lx exhibited higher dynamic binding capacities than mabselect sure at longer residence times, while at shorter residence times they.
Example of hiscreen mabselect prisma hitrap and hiscreen mabselect prisma offer the highest dbc compared to other ges protein a columns mabselect prisma mabselect sure lx mabselect sure ka3580210418pp 17 hiscreen mabselect prisma dbc compared to hiscreen mabselect sure lx and hiscreen mabselect sure at different residence time. For all other nonv h 3 antibody subfamilies tested i. Mabselect sure lx is an affinity chromatography medium for capturing high titer monoclonal antibodies from large volumes of feed by packed. We specialize in automated highthroughput protein and. Mabselect sure lx is an alkalistabilized, protein aderived affinity medium with a binding capacity for monoclonal antibodies mabs exceeding that of. Mabselect is designed for highthroughput purification of monoclonal antibodies from large volumes of feed. Evaluation of a nextgeneration protein a chromatography. Sure lx, and the prototype mabselect prisma, respectively. Because of the elevated capacities observed for the. This section describes purification of monoclonal antibodies with hitrap mabselect sure. Mabselect is a bioprocess affinity medium for capturing monoclonal antibodies from large volumes of feed by packed bed chromatography.
Mabselect sure lx 70 40 60 30 10 50 20 0 0 2 4 6 8 10 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 0,0 0 50 100 150 200 250 300 350 400 y0,0048x r20,9884 linear flow velocity cmh pressure flow bar dynamic binding capacity after cleaning in place 0. Characteristics of mabselect sure lx 1 d 50v is the medium particle size of the cumulative volume distribution. Mabselect prisma mabselect sure lx mabselect sure 0 10 20 30 40 50 60 70 80 90 100 1 25 50 100 125 150 cycle mabselect prisma remaining relative dbc q mabselect sure lx b10, % mabselect prisma exhibits significantly increased dynamic binding capacity compared with its predecessors. The performance of mabselect sure lx, an alkalistabilized, protein aderived affinity medium resin for capturing monoclonal antibodies, was evaluated over 100 purification cycles that included cleaninginplace with 0.
Mabselect prisma mabselect sure lx mabselect sure 0 20 40 60 0 25 50 75 100 125 150 relative capacity, qb10 % cycle, 1. Mabselect sure resin offers generic elution conditions for different monoclonal antibodies, which is an advantage when designing purification platform processes. Data file 18114994 ae affinity chromatography mabselect. The results obtained for mabselect are shown in figure 2. Data file 11001165 ac affinity chromatography mabselect. Captiva primab exhibited comparable dbc 100 at rt 2 min as the resin with lowest dbc 100 at prolonged rts, poros. Mabselect sure lx and mabselect sure have the same particle size and pore size, while mabselect sure lx has higher ligand densities as claimed by the manufacturer. Mabselect xtra demonstrates even higher dbc and is also optimized for fcfusion proteins. Assay description the assay is intended for quantification of protein a ligands in relation to a defined concentration of igg. The prototype mabselect prisma dbc also was 50% higher in dbc at the lowest residence time of 2 minutes, whereas mabselect sure and mabselect sure lx appear to converge to the same value.
In continuous chromatography, however, the lower flow velocity of mabselect sure pcc is counterbalanced by the use of smaller columns with shorter bed heights. These high capacity stationary phases represent a diverse mix of properties likely to impact process performance, including mass transfer rates 1. Columns and resins for antibody purification and immunoprecipitation. Lwell 4 plates 28925825 mabselect sure lx same features as mabselect sure with increased antibody binding capacity. Optimized crosslinking of the agarose base matrix allows for, at minimum, five fold. Improved proteina separation of vh3 fab from fc after. Healthcares mabselect family is called mabselect sure lx 2. Mabselect sure lx 2015 mabselect sure pcc increased mass throughput and bioburden control 2017 mabselect prisma 2001 mabselect 2005 mabselect sure 1950 1980 2000 2020 evolution of ge healthcares protein a chromato. Sure lx, an alkalistabilized, protein aderived affinity medium resin for capturing monoclonal antibodies. Frontal analysis was performed with native and spiked feeds. Linear v elocit cm 700 600 500 400 300 200 100 0 0 0. Lwell 4 plates 28925824 predictor mabselect sure, 50. The associated increase in contaminant levels, such as host cell proteins hcp, viruses, aggregates, and protein a, leads to new challenges for mab purification, and upgraded processes with efficient protein purification steps are required. Ge healthcare mabselect sure lx chromatography media protein a affinity medium with high dynamic binding capacity which improves performance and cuts costs manufacturer.
Hitrap mabselect sure hitrap mabselect hitrap mabselect xtra. Performing a purification of igg antibodies with mabselect. Instructions29262586ac affinitychromatography mabselect. Relative remaining dynamic binding capacity using 1. A strategy for process dev elopment using multicolumn. Mabselect sure is composed of the same rigid, high flow agarose matrix, but is based on an alkalistabilized rprotein abased ligand. In addition, the recombinant protein a ligand of this resin is engineered to include a. Lifetime performance study of mabselect sure lx during. Mabselect, mabselect sure, and mabselect sure lx mabselect resin is a recombinant protein abased affinity resin. The optimized crosslinked agarose matrix of the resin gives at least five times higher fluid velocities compared with conventional crosslinking. Phytip proplus affinity columns the best of protein a and. Ge healthcare hiscreen mabselect, xtra, sure, sure lx. Data file 18114994 af affinity chromatography mabselect. Use of benzyl alcohol as a shipping and storage solution.
This feature is also correlated to the construction of the rprotein a ligand, which has a reduced interaction with the vh3 fab domain present in human antibodies. Mabselect sure lx protein a resin cytiva, formerly ge healthcare. For more information on mabselect and mabselect xtra, refer to data files 18114994 and 11001157 respectively. Clarified cho cell culture supernatant was applied to the column until approximately 10% breakthrough was achieved. Hitrap mabselect sure, hitrap mabselect, and hitrap mabselect xtra 1 and 5 ml for purification of monoclonal antibodies with optimized binding capacities at high flow rates. Sure lx to meet the needs associated with hightiter upstream processes. Serial dilution guidelines prepare serial dilutions using the prediluted.
As an example, at 6 min residence time, the dynamic binding capacity of mabselect sure lx for human igg is approximately 60 gl. As table 1 shows, it has been developed using the same highly crosslinked agarose base matrix and protein a ligand as for other mab affinity resins table 1. Hitrap mabselect sure 5 ml, 1 x 5 ml 11003494 store between the following temperatures. Ge healthcare 28401860, see predilute the ge healthcare mabselect sure protein a standard on page 11.
1349 337 1043 735 303 1340 367 203 1289 1512 1270 1534 863 575 887 334 575 624 510 1312 228 593 643 1528 1385 255 402 311 1374 951 323 1516 744 152 979 249 763 855 1173 1371 4 345 1093 473 353 1331 866