Downstream Process Development and Tech Transfer of novel polyclonal antibody therapeutical products

• Initiated optimization of a PhIII mAb downstream process and demonstrated addition of an Emphaze filter to the harvest operation was beneficial for downstream processes.• Supported analytical team’s late-stage development activities by developing methods to isolate populations of product variants, such as glycated forms.• Used Surrogate MockV-MVM kit to investigate the restoration of Viral Clearance of a mAb AIEX step. • Supported regulatory submission activities such as process mapping, change control and risk assessments..• Developed protein purification methods for clinical candidate lead molecules.• Performed material supply runs for formulation development.

• Developed and established a challenging capturing chromatography for a cytokine drug candidate, including resin selection and method development. Used Robocolumns to screen binding and elution on Tecan, verified and consolidated method on AKTA FPLC using small, middle sized columns. Worked with other downstream SMEs all the way to generate GLP/Tox material at 200L scale. • Chromatography development for a bispecific antibody: screening of conditions of MabSelect Sure capture step; established a HIC FT mode intermediate step to remove >10% aggregates after screening more than a dozen of resins, optimized the HIC FT condition using DOE on Unicorn and JMP for analysis; developed an alternative Mixed mode polishing step using Isocratic elution; in charge of large scale intermediate and polishing step PD runs • The processes were successfully transferred to manufacture for GMP• Developed polishing step chromatography for 2 target antibodies, tested in confirmatory runs at bench and pilot scales• The processes were successfully transferred to manufacture GLP/GMP runs • Authorized and reviewed development reports and tech transfer reports, participated and completed tech transfers to manufacture • Reviewed and edited master control documents such as MBRs and SOPs for manufacture• On-site support for manufacture during GLP and GMP runs as SME• Trouble shooting and scale-down experiments for deviation of process scaleup to manufacture• Tested changes of existing processes, or add-on new process step to mitigate risk, and transferred to manufacture for GMP runs • Material runs to provide study materials for DSP and AD, including low endo batches for preclinical Tox studies

• Provided regular CRO down-stream bio-production services, including services strictly following manufacture batch record. From cell harvesting, protein purification on AKTA or drip columns, formulation through TFF or concentration & dialysis.• Purified antibodies, antibody fragments, and biologics using Affinity, IEX, HIC, and Gel Filtration techniques. Purification method development and optimization. Proficient in Unicorn software and familiar on different lab scale AKTAs.• Performed downstream process development pilot studies and scale-ups for clients. Successfully transferred batch records and SOPs to CMOs for clients.• Protein formulation and stability test.• Performed protein analytic work including: LAL, SEC-HPLC, CIEX-HPLC, RP-HPLC, Normal-HPLC, N-glycan analysis, IEF, Western Blot, SDS-PAGE, and ELISA.• Good knowledge of other analytic techniques such as SEC-MALS, DLS, DSC, AUC, CE-SDS and CIEF etc.• Wrote reports and summaries and interacted with clients regularly

• Optimized 5L bioreactor runs of 2 mAbs expressed in CHO cells via screening or titration of: cell lines; seeding density; nutrient feeding schedule; pH and DO value; gas-flow control; impeller type and agitation speed; sparger type. Final IgG production levels reached 1.5 g/L, and 2 g/L, respectively.• Sub-cloned of mAb GS constructs in CHO-S Cells; picked colonies and performed 48-hr-production test; further screened top clones in 5 ml and 25 ml-production tests• Performed nutrient-feeding tests for different cell lines in shaker tubes and flasks• Provided an internal Protein A IgG-quantitative HPLC daily service and a regular mAb variants-evaluation CEX HPLC service• Trained new employees and interns lab-techniques and to perform routine internal services• Adapted CHO-K1 into a serum-free suspension cell line

• Assay support, including HPLC for t-PA/TNK, Affinity and Reverse Phase Analysis of antibody fragments, Nutropin Reversed Phase HPLC, and HTS quantification of IgG in CHO HCCF via protein A HPLC• Coordinated work with others in the department to release quality results within the timeline specified for a given assay • General lab maintenance • Followed written procedures and protocols demonstrating attention to detail

• Antibody production, purification and analysis of ~100 mouse, chimera, and human monoclonal antibodies. • Optimized a method for generating Fab fragments for human IgG3 molecules. Used Fab fragments and whole antibody to determine binding constants for antibody: antibody interactions• Determined immunoglobulin quantity for human, mouse, and chimera immunoglobulin molecules using the ForteBio Octet System• Developed a conjugation method for antibodies otherwise difficult to conjugate• Optimized antibody production levels• Tested multiple formulations to optimize solubility of purified antibody• Trained associates in experimental procedures and instrument operation