Preclinical immunogenicity testing using anti-drug antibody analysis of GX-G3, Fc-fused recombinant human granulocyte colony-stimulating factor, in rat and monkey models

• Published in: Scientific reports Vol. 11; no. 1; p. 12004
• Main Authors: Kim, Yun Jung, Koh, Eun Mi, Song, Chi Hun, Byun, Mi Sun, Choi, Yu Ri, Jeon, Eun-Jeong, Hwang, Kyunghwa, Kim, Sang Kyum, Yang, Sang In, Jung, Kyung Jin
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07-06-2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/664/2228; 631/250/249; 692/499; Acute myeloid leukemia; Animal models; Antibodies; Cell proliferation; Chemotherapy; Colonies; Colony-stimulating factor; Granulocyte colony-stimulating factor; Granulocytes; Humanities and Social Sciences; Immunogenicity; Leukemia; Leukocytes (granulocytic); Malignancy; multidisciplinary; Myeloid leukemia; Neutropenia; Science; Science (multidisciplinary); Serum levels; Toxicity testing
• DOI: 10.1038/s41598-021-91360-7
• EISSN: 2045-2322

Human granulocyte colony-stimulating factor (G-CSF, this study used Fc-fused recombinant G-CSF; GX-G3) is an important glycoprotein that stimulates the proliferation of granulocytes and white blood cells. Thus, G-CSF treatment has been considered as a crucial regimen to accelerate recovery from chemotherapy-induced neutropenia in cancer patients suffering from non-myeloid malignancy or acute myeloid leukemia. Despite the therapeutic advantages of G-CSF treatment, an assessment of its immunogenicity must be performed to determine whether the production of anti-G-CSF antibodies causes immune-related disorders. We optimized and validated analytical tools by adopting validation parameters for immunogenicity assessment. Using these validated tools, we analyzed serum samples from rats and monkeys injected subcutaneously with GX-G3 (1, 3 or 10 mg/kg once a week for 4 weeks followed by a 4-week recovery period) to determine immunogenicity response and toxicokinetic parameters with serum concentration of GX-G3. Several rats and monkeys were determined to be positive for anti-GX-G3 antibodies. Moreover, the immunogenicity response of GX-G3 was lower in monkeys than in rats, which was relevant to show less inhibition of toxicokinetic profiles in monkeys, at least 1 mg/kg administrated group, compared to rats. These results suggested the establishment and validation for analyzing anti-GX-G3 antibodies and measurement of serum levels of GX-G3 and anti-GX-G3 antibodies, which was related with toxicokinetic profiles. Taken together, this study provides immunogenicity assessment which is closely implicated with toxicokinetic study of GX-G3 in 4-week repeated administrated toxicological studies.