Mabselect sure lx pdf

Because of the elevated capacities observed for the. Mabselect sure is composed of the same rigid, high flow agarose matrix, but is based on an alkalistabilized rprotein abased ligand. Performing a purification of igg antibodies with mabselect. Data file 11001165 ac affinity chromatography mabselect. The latest member of ge healthcares mabselect family is called mabselect sure lx. Mabselect sure lx is an affinity chromatography medium for capturing high titer monoclonal antibodies from large volumes of feed by packed. Improved proteina separation of vh3 fab from fc after. One of the key features of mabselect sure lx is its increased dynamic binding. Mabselect, mabselect sure, and mabselect sure lx mabselect resin is a recombinant protein abased affinity resin. Serial dilution guidelines prepare serial dilutions using the prediluted. Hitrap mabselect sure 5 ml, 1 x 5 ml 11003494 store between the following temperatures. For all other nonv h 3 antibody subfamilies tested i.

Mabselect sure lx and mabselect sure were packed in tricorn 5100 columns bed height 10 cm, column volume 1. Sure lx to meet the needs associated with hightiter upstream processes. Instructions29262586ac affinitychromatography mabselect. Mabselect sure resin offers generic elution conditions for different monoclonal antibodies, which is an advantage when designing purification platform processes. Linear v elocit cm 700 600 500 400 300 200 100 0 0 0. 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. Data file 18114994 af affinity chromatography mabselect. 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 prisma mabselect sure lx mabselect sure 0 20 40 60 0 25 50 75 100 125 150 relative capacity, qb10 % cycle, 1. 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. 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. Use of benzyl alcohol as a shipping and storage solution. Lwell 4 plates 28925824 predictor mabselect sure, 50. Plate number, assymetry and pressure vs flow were determined.

As a result, mabselect sure lx exhibited higher dynamic binding capacities than mabselect sure at longer residence times, while at shorter residence times they. In continuous chromatography, however, the lower flow velocity of mabselect sure pcc is counterbalanced by the use of smaller columns with shorter bed heights. The mabselect sure ligand was developed by protein engineering of one of the iggbinding domains of protein a. 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. Clarified cho cell culture supernatant was applied to the column until approximately 10% breakthrough was achieved. Phytip proplus affinity columns the best of protein a and.

Assay description the assay is intended for quantification of protein a ligands in relation to a defined concentration of igg. Hitrap mabselect sure hitrap mabselect hitrap mabselect xtra. Sure lx, an alkalistabilized, protein aderived affinity medium resin for capturing monoclonal antibodies. 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. Mabselect prisma in comparison with mabselect sure and mabselect sure lx. Ge healthcare hiscreen mabselect, xtra, sure, sure lx. We specialize in automated highthroughput protein and.

Lwell 4 plates 28925825 mabselect sure lx same features as mabselect sure with increased antibody binding capacity. 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. Healthcares mabselect family is called mabselect sure lx 2. Typical pressureflow profile for mabselect sure in a packed bed bed height 20 cm in a bpg 300 column i. 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. The breakthrough capacity q b, 10% and additional stability indicating parameters were measured at 0, 6, 12, 24, and 36 months. Captiva primab exhibited comparable dbc 100 at rt 2 min as the resin with lowest dbc 100 at prolonged rts, poros. Here, a fixed column size of 450200 mm, a mab titer of 4 gl, and a 20% safety factor in loading were used. In addition, the recombinant protein a ligand of this resin is engineered to include a. Results showed that dynamic binding capacity was maintained and that yield was constant throughout the study. Ge healthcare 28401860, see predilute the ge healthcare mabselect sure protein a standard on page 11. Data file 18114994 ae affinity chromatography mabselect fig 1.

Application note 28907892 ad processscale antibody. Characteristics of mabselect sure lx 1 d 50v is the medium particle size of the cumulative volume distribution. Ge healthcare mabselect sure lx chromatography media protein a affinity medium with high dynamic binding capacity which improves performance and cuts costs manufacturer. 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. As an example, at 6 min residence time, the dynamic binding capacity of mabselect sure lx for human igg is approximately 60 gl. Sure lx, and the prototype mabselect prisma, respectively. 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. Mabselect is designed for highthroughput purification of monoclonal antibodies from large volumes of feed. Evaluation of a nextgeneration protein a chromatography. Protein a affinity chromatography of chinese hamster ovary. 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.

Relativedynamicbindingcapacity,qb10%,formabselectprismaafter cipwith0. Mabselect xtra demonstrates even higher dbc and is also optimized for fcfusion proteins. Lifetime performance study of mabselect sure lx during. The results obtained for mabselect are shown in figure 2. Toyopearl afrprotein a hc650f toyopearl afrprotein a650f mabselect sure lx column size. Data file 11003490 ac affinity chromatography hitrap mabselect sure hitrap mabselect hitrap mabselect xtra fig 1. Data file 18114994 ae affinity chromatography mabselect. Frontal analysis was performed with native and spiked feeds. Mabselect prisma affinity columns and resins for antibody. Data file 11003490 ac affinity chromatography hitrap. Combining the proven technology of phytip affinity columns with the benefits of a new resin demonstrating attributes of both protein a and. Mabselect protein a resins ge healthcare life sciences. Dynamic binding capacity study on mabselect sure lx for.

Protein a quantification kit, see predilute the repligen protein a standard on page 11. Packing mabselect and mabselect sure resins using verified. Relative remaining dynamic binding capacity using 1. For more information on mabselect and mabselect xtra, refer to data files 18114994 and 11001157 respectively. Ge healthcare mabselect sure lx chromatography media pack. Mabselect sure lx protein a resin cytiva, formerly ge healthcare. These high capacity stationary phases represent a diverse mix of properties likely to impact process performance, including mass transfer rates 1. The optimized crosslinked agarose matrix of the resin gives at least five times higher fluid velocities compared with conventional crosslinking. Here, we provide an overview of a novel system for mab capture from a users perspective. 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.

Mabselect sure lx has been further developed from mabselect sure to. Mabselect sure lx is a protein a affinity resin with very high dynamic binding capacity at extended residence times. Mabselect sure lx is developed for hightiter mab capture at process scale. This section describes purification of monoclonal antibodies with hitrap mabselect sure. The ligand provides greater stability than conventional rprotein abased media, allowing extended use of 0. Mabselect sure lx is an alkalistabilized, protein aderived affinity medium with a binding capacity for monoclonal antibodies mabs exceeding that of. Optimized crosslinking of the agarose base matrix allows for, at minimum, five fold. Mabselect sure lx mabselect sure lx is an alkalistabilized, protein aderived affinity medium with a binding capacity for monoclonal antibodies mabs exceeding that of mabselect sure at longer residence times. A strategy for process dev elopment using multicolumn. Mabselect is a bioprocess affinity medium for capturing monoclonal antibodies from large volumes of feed by packed bed chromatography. 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. Columns and resins for antibody purification and immunoprecipitation. 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.