GMP COMPLIANT MANUFACTURING PROCESS DEVELOPMENT OF ONCOLYTIC VIROTHERAPY

Background & Aim Development of oncolytic virotherapy for cancer treatment is attracting great concerns as a next-generation innovative modality. Oncolytic virotherapy usually utilize genetically modified virus that is designed to multiply only in cancer cells for safety. Conducting clinical trials as well as pre-clinical studies requires large quantities of high quality viruses as to GMP. Here we present the key considerations regarding GMP compliant process development of manufacturing oncolytic virus, Coxsackie virus type B3 (CVB3), designed for clinical trial targeting triple-negative breast cancer. CVB3 is a non-enveloped, linear single-strand RNA virus, and sizing approximately 27 to 33 nm diameter. Methods, Results & Conclusion Here we will discuss the key considerations we have gained during the manufacturing process development from the initial model using a zonal rotor centrifugal separator to the later developed systems including tangential flow filtration system and ion chromatography. Later developed system consists of all process, from upstream cell culture expansion to downstream target purification, being designed as fully closed and single-use manufacturing system. In brief, HEK293 cell suspension extended in 3L serum-free medium infected with CVB3, made total 150 mL of final products as 8.43 x 10 exp7 TCID50/mL concentration. Quality analysis of products related impurities as residual human cell protein and remnant DNA/RNA was also confirmed to be within FDA recommended standards. Thus the GMP compliant manufacturing process of oncolytic virus we established considered to be suitable for technology transfer for future commercial production. Development of oncolytic virotherapy for cancer treatment is attracting great concerns as a next-generation innovative modality. Oncolytic virotherapy usually utilize genetically modified virus that is designed to multiply only in cancer cells for safety. Conducting clinical trials as well as pre-clinical studies requires large quantities of high quality viruses as to GMP. Here we present the key considerations regarding GMP compliant process development of manufacturing oncolytic virus, Coxsackie virus type B3 (CVB3), designed for clinical trial targeting triple-negative breast cancer. CVB3 is a non-enveloped, linear single-strand RNA virus, and sizing approximately 27 to 33 nm diameter. Here we will discuss the key considerations we have gained during the manufacturing process development from the initial model using a zonal rotor centrifugal separator to the later developed systems including tangential flow filtration system and ion chromatography. Later developed system consists of all process, from upstream cell culture expansion to downstream target purification, being designed as fully closed and single-use manufacturing system. In brief, HEK293 cell suspension extended in 3L serum-free medium infected with CVB3, made total 150 mL of final products as 8.43 x 10 exp7 TCID50/mL concentration. Quality analysis of products related impurities as residual human cell protein and remnant DNA/RNA was also confirmed to be within FDA recommended standards. Thus the GMP compliant manufacturing process of oncolytic virus we established considered to be suitable for technology transfer for future commercial production.