이중항체 의약품의 연구개발 동향

Sun-Il Kwon 대한의생명과학회 2020 Biomedical Science Letters Vol.26 No.3

A bispecific antibody (BsAb) is an artificial protein containing two kinds of specific antigen binding sites. BsAb can connect target cells to functional cells or molecules, and thus stimulate a directed immune response. Last several decades a wide variety of bsAb formats and production technologies have been developed. BsAbs are constructed either chemically or biologically, exploiting techniques like cell fusion and recombinant DNA technologies. There are over 100 different formats of bsAb so far developed, but they could be classified into the two main categories such as Fc-based (with a Fc region) bsAbs and fragment-based (without a Fc region) bsAbs. BsAb has a broad application prospect in tumor immunotherapy and drug delivery. Here, we present a brief introduction to the structure of antibody, pharmacological mechanisms of antibodies and the trend in the production technologies of therapeutic antibodies. In addition, we address a review on the current status of various bsAb format development and their production technologies together with global situation in the clinical studies of bsAb.

Purification and characterization of monoclonal IgG antibodies recognizing Ebola virus glycoprotein

한백상,장호영,Trina Racine,Xiangguo Qiu,신정임 대한백신학회 2018 Clinical and Experimental Vaccine Research Vol.7 No.2

Purpose: The goal of this study was to purify and characterize Ebola virus glycoprotein (GP)-specific IgG antibodies from hybridoma clones. Materials and Methods: For hybridoma production, mice were injected by intramuscular-electroporation with GP DNA vaccines, and boosted with GP vaccines. The spleen cells were used for producing GP-specific hybridoma. Enzyme-linked immunosorbent assay, Western blot assay, flow cytometry, and virus-neutralizing assay were used to test the ability of monoclonal IgG antibodies to recognize GP and neutralize Ebola virus. Results: Twelve hybridomas, the cell supernatants of which displayed GP-binding activity by enzyme-linked immunosorbent assay and the presence of both IgG heavy and light chains by Western blot assay, were chosen as a possible IgG producer. Among these, five clones (C36-1, D11-3, D12-1, D34-2, and E140-2) were identified to secrete monoclonal IgG antibodies. When the monoclonal IgG antibodies from the 5 clones were tested for their antigen specificity, they recognized GP in an antigen-specific and IgG dose-dependent manner. They remained reactive to GP at the lowest tested concentrations (1.953-7.8 ng/mL). In particular, IgG antibodies from clones D11-3, D12-1, and E140-2 recognized the native forms of GP expressed on the cell surface. These antibodies were identified as IgG1, IgG2a, or IgG2b kappa types and appeared to recognize the native forms of GP, but not the denatured forms of GP, as determined by Western blot assay. Despite their GP-binding activity, none of the IgG antibodies neutralized Ebola virus infection in vitro, suggesting that these antibodies are unable to neutralize Ebola virus infection. Conclusion: This study shows that the purified IgG antibodies from 5 clones (C36-1, D11-3, D12-1, D34-2, and E140-2) possess GP-binding activity but not Ebola virus-neutralizing activity.

Antibody response to COVID-19 vaccination in patients on chronic hemodialysis

박인휘 대한백신학회 2023 Clinical and Experimental Vaccine Research Vol.12 No.3

Purpose: Since patients on hemodialysis (HD) are known to be vulnerable to coronavirus disease 2019 (COVID-19), many studies were conducted regarding the effectiveness of the COVID-19 vaccine in HD patients in Western countries. Here, we assessed antibody response of HD patients for 6 months post-vaccination to identify the duration and effectiveness of the COVID-19 vaccine in the Asian population. Materials and Methods: We compared antibody response of the COVID-19 vaccine in HD patients with healthy volunteers. Patient and control groups had two doses of ChAdOx1 nCoV-19 and mRNA-1273, respectively. Immunoglobulin G (IgG) was measured before vaccination, 2 weeks after the first dose, 2 and 4 weeks, 3 and 6 months after the second dose. Neutralizing antibody was measured before vaccination and at 2 weeks, 3 and 6 months after second dose. Since the third dose was started in the middle of the study, we analyzed the effect of the third dose as well. Results: Although antibody production was weaker than the control group (n=22), the patient group (n=39) showed an increase in IgG and neutralizing antibody after two doses. And, 21/39 patients and 14/22 participants had a third dose (BNT162b2 or mRNA-1273 in the patient group, mRNA-1273 in the control group), and it did not affect antibody response in both group. Trend analysis showed IgG and neutralizing antibody did not decrease over time. Age, sex, and HD vintage did not affect antibody production in HD patients. Patients with higher body mass index displayed better seroresponse, while those on immunosuppressants showed poor seroresponse. Conclusion: Two doses of vaccination led to significant antibody response in HD patients, and the antibody did not wane until 6 months.

Bone Generation Following Repeated Administration of Recombinant Bone Morphogenetic Protein 2

Son Hye-Ju,Lee Mi Nam,Kim Yuri,Choi Hyuck,Jeong Byung-Chul,Oh Sin-Hye,김정우,Kwon Seung-Hee,Kim Sun-Hun,Song Soo-Chang,Lee Shee Eun,Koh Jeong-Tae 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.1

Background: The delivery of recombinant human bone morphogenetic protein 2 (rhBMP2) by using various carriers has been used to successfully induce bone formation in many animal models. However, the effect of multiple administration of rhBMP2 on bone formation and BMP2 antibody production has not been determined. Our aim was to examine the bone formation activity of rhBMP2 and serum levels of anti-BMP2 antibodies following the repeated administration of rhBMP2 in mice. Methods: Absorbable collagen sponges or polyphosphazene hydrogels containing rhBMP2 were subcutaneously implanted or injected into one side on the back of six-week-old C57BL/6 mice. Three or 4 weeks later, the same amount of rhBMP2 was administered again with the same carrier into the subcutaneous regions on the other side of the back or into calvarial defects. The effects of a single administration of rhBMP2 on the osteoinductive ability in the ectopic model were compared with those of repeated administrations. In vivo ectopic or orthotopic bone formation was evaluated using microradiography and histological analyses. Serum concentrations of anti-rhBMP2 antibodies were measured by ELISAs. Results: Re-administration of the same amount of rhBMP2 into the subcutaneous area showed a comparable production of ectopic bone as after the first administration. The bone forming ability of repeated rhBMP2 administrations was equal to that of single rhBMP2 administration. The administration of rhBMP2 into calvarial defects, following the first subcutaneous administration of rhBMP2 on the back, completely recovered the defect area with newly regenerated bone within 3 weeks. Repeated administration of rhBMP2 at 4-week intervals did not significantly alter the serum levels of anti-BMP2 antibodies and did not induce any inflammatory response. The serum obtained from rhBMP2-exposed mice had no effect on the ability of rhBMP2 to induce osteogenic gene expressions in MC3T3-E1. Conclusion: We suggest that the osteoinductive ability of rhBMP2 is not compromised by repeated administrations. Thus, rhBMP2 can be repeatedly used for bone regeneration at various sites within a short duration. Background: The delivery of recombinant human bone morphogenetic protein 2 (rhBMP2) by using various carriers has been used to successfully induce bone formation in many animal models. However, the effect of multiple administration of rhBMP2 on bone formation and BMP2 antibody production has not been determined. Our aim was to examine the bone formation activity of rhBMP2 and serum levels of anti-BMP2 antibodies following the repeated administration of rhBMP2 in mice. Methods: Absorbable collagen sponges or polyphosphazene hydrogels containing rhBMP2 were subcutaneously implanted or injected into one side on the back of six-week-old C57BL/6 mice. Three or 4 weeks later, the same amount of rhBMP2 was administered again with the same carrier into the subcutaneous regions on the other side of the back or into calvarial defects. The effects of a single administration of rhBMP2 on the osteoinductive ability in the ectopic model were compared with those of repeated administrations. In vivo ectopic or orthotopic bone formation was evaluated using microradiography and histological analyses. Serum concentrations of anti-rhBMP2 antibodies were measured by ELISAs. Results: Re-administration of the same amount of rhBMP2 into the subcutaneous area showed a comparable production of ectopic bone as after the first administration. The bone forming ability of repeated rhBMP2 administrations was equal to that of single rhBMP2 administration. The administration of rhBMP2 into calvarial defects, following the first subcutaneous administration of rhBMP2 on the back, completely recovered the defect area with newly regenerated bone within 3 weeks. Repeated administration of rhBMP2 at 4-week intervals did not significantly alter the serum levels of anti-BMP2 antibodies and did not induce any inflammatory response. The serum obtained from rhBMP2-exposed mice had no effect on the ability of rhBMP2 to induce osteogenic gene expressions in MC3T3-E1. Conclusion: We suggest that the osteoinductive ability of rhBMP2 is not compromised by repeated administrations. Thus, rhBMP2 can be repeatedly used for bone regeneration at various sites within a short duration.

Positivity of SARS-CoV-2 Antibodies among Korean Healthy Healthcare Workers 1 and 2 Weeks after Second Dose of Pfizer-BioNTech Vaccination

Kim Namhee,Minn Dohsik,Park Seungman,Roh Eun Youn,Yoon Jong Hyun,Park Hyunwoong,Shin Sue 대한의학회 2021 Journal of Korean medical science Vol.36 No.21

The antibody titer of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was observed in 289 healthy healthcare workers who had completed the second dose of the Pfizer-BioNTech coronavirus disease 2019 (COVID-19) vaccine. Antibody tests were performed using both the automated electrochemiluminescence immunoassay (ECLIA) and the chromatographic lateral flow immunoassay (LFIA). All subjects had antibodies against the receptor binding domain of the spike protein of SARS-CoV-2 only one week after completing the vaccination, and the antibody titer became significantly higher after another week (P < 0.001). Since there was a large amount of antibody formation within two weeks after completion of vaccination, the less sensitive method, LFIA, also showed high sensitivity. There was no significant difference between whole blood and serum in detecting SARS-CoV-2 antibodies after vaccination. This is an early study of vaccinations among Koreans and is expected to contribute to the establishment of national guidelines on COVID-19 vaccination.

Comparison of the rapidity of SARS-CoV-2 immune responses between primary and booster vaccination for COVID-19

Ji Yeun Kim,Ji-Soo Kwon,Hye Hee Cha,So Yun Lim,Seongman Bae,Sung-Han Kim 대한내과학회 2022 The Korean Journal of Internal Medicine Vol.37 No.6

Background/Aims: The rapidity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific memory B or T cell response in vaccinated individuals is important for our understanding of immunopathogenesis of coronavirus disease 2019 (COVID-19). We therefore compared the timing of adequate immune responses between the first and booster doses of COVID-19 vaccines in infection-naïve healthcare workers. Methods: We enrolled healthcare workers who received two doses of either the BNT162b2 vaccine or the ChAdOx1 vaccine, all of whom received the BNT162b2 vaccine as the booster (the third) dose. Spike 1 (S1)-immunoglobulin G (IgG) antibodies and interferon gamma producing T cell responses were measured at 0, 7, 14, and 21 days after the first dose, and at 0 and between 2 to 7 days after the booster dose. Results: After the first-dose vaccination, the S1-IgG antibody responses were elicited within 14 days in the BNT162b2 group and within 21 days in the ChAdOx1 group. After the booster dose, the S1-IgG antibody responses were elicited within 5 days in both groups. The SARS-CoV-2-specific T cell responses appeared at 7 days after the first dose and at 4 days after the booster dose. Conclusions: SARS-CoV-2-specific immune responses by memory B cells and T cells may be expected to appear around 4 to 5 days after the booster dose.

Humoral response to viral vector COVID-19 vaccine in hemodialysis patients

Raja Niranjan,Rajagopalan Arul,Arunachalam Jegan,Prasath Arun,Durai Rakesh,Rajendran Manorajan 대한신장학회 2022 Kidney Research and Clinical Practice Vol.41 No.3

Background: The coronavirus disease 2019 (COVID-19) vaccine is not readily available in many countries where dosing interval is spaced more than ideal. Patients with chronic kidney disease, especially those on maintenance hemodialysis, have a tendency for a reduced immune response. This study was undertaken to demonstrate the distinct humoral immune response to the viral vector COVID-19 vaccine in patients with kidney failure receiving maintenance hemodialysis. Methods: The study was carried out with two cohorts: 1) patients receiving maintenance hemodialysis and 2) healthcare workers from the same dialysis center as controls, each group with 72 subjects. Participants received a dose of Covishield ChAdOx1 nCoV-19 coronavirus vaccine. The humoral immunological response was determined using electrochemiluminescence immunoassay which quantitatively measures antibodies to the severe acute respiratory syndrome coronavirus 2 spike protein receptor-binding domain. Results: All study subjects in the control group developed a humoral response (antibody titer of ≥0.8 U/mL), while only 64 of 72 in the dialysis group (88.9%) were responders. Age (ρ = –0.234, p = 0.04) and sodium level (ρ = 0.237, p = 0.04) correlated with low antibody titer in bivariate analysis. In multivariate analysis, only age (odds ratio, 1.10; 95% confidence interval, 1.01–1.22; p = 0.045) was associated with nonresponders. Conclusion: Our study demonstrated a weak antibody response of hemodialysis patients to the viral vector COVID-19 vaccine. Older age was associated with nonresponders. Evaluation of both humoral and cellular immunity after the second vaccine dose and serial antibody titers can help determine the need for booster shots.

Circulating Anti-Elastin Antibody Levels and Arterial Disease Characteristics: Associations with Arterial Stiffness and Atherosclerosis

이승현,이상학,신기혁,박성하,강석민,최동훈,이승효 연세대학교의과대학 2015 Yonsei medical journal Vol.56 No.6

Purpose: Elastin is a major arterial structural protein, and elastin-derived peptides are related to arterial change. We previously reported on a novel assay developed using aortic elastin peptides; however, its clinical implications remain unclear. In this study, we assessed whether anti-elastin antibody titers reflect the risk of coronary artery disease (CAD) or its characteristics. Materials and Methods: We included 174 CAD patients and 171 age- and sex-matched controls. Anti-elastin antibody titers were quantified by enzyme-linked immunosorbent assay. Parameters of arterial stiffness, including the augmentation index (AI) and heart-to-femoral pulse wave velocity (hfPWV), were measured non-invasively. The clinical and angiographic characteristics of CAD patients were also evaluated. Associations between anti-elastin levels and vascular characteristics were examined by linear regression analysis. Results: The median blood level of anti-elastin was significantly lower in the CAD group than in the controls [197 arbitrary unit (a.u.) vs. 63 a.u., p<0.001]. Levels of anti-elastin were significantly lower in men and in subjects with hypertension, diabetes mellitus,hyperlipidemia, or high hfPWV. Nevertheless, anti-elastin levels were not dependent on atherothrombotic events or the angiographicseverity of CAD. In a multivariate analysis, male sex (β=-0.38, p<0.001), diabetes mellitus (β=-0.62, p<0.001), hyperlipidemia(β=-0.29, p<0.001), and AI (β=-0.006, p=0.02) were ultimately identified as determinants of anti-elastin levels. Conclusion: Lower levels of anti-elastin are related to CAD. The association between antibody titers and CAD is linked to arterial stiffness rather than the advancement of atherosclerosis.

Improving vaccines by targeting antigens to dendritic cells

Ken Shortman,Mireille H. Lahoud,Irina Caminschi 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.2

A new approach to enhancing the effectiveness of vaccines is to deliver antigens selectively to dendritic cells (DC) in situ, via monoclonal antibodies specific for particular DC surface molecules. This can markedly enhance CTL responses and, via helper T cells, also enhance antibody responses. DC activation agents or adjuvants must also be administered for effective CTL responses, but in some cases good antibody responses can be obtained without adjuvants. Here we review the role of different DC subsets and different DC target molecules in obtaining enhanced immune responses. A new approach to enhancing the effectiveness of vaccines is to deliver antigens selectively to dendritic cells (DC) in situ, via monoclonal antibodies specific for particular DC surface molecules. This can markedly enhance CTL responses and, via helper T cells, also enhance antibody responses. DC activation agents or adjuvants must also be administered for effective CTL responses, but in some cases good antibody responses can be obtained without adjuvants. Here we review the role of different DC subsets and different DC target molecules in obtaining enhanced immune responses.

Comparison of antibody responses after the 1st and 2nd doses of COVID-19 vaccine with those of patients with mild or severe COVID-19

( Hye Hee Cha ),( So Yun Lim ),( Ji-soo Kwon ),( Ji Yeun Kim ),( Seongman Bae ),( Jiwon Jung ),( Sung-han Kim ) 대한내과학회 2022 The Korean Journal of Internal Medicine Vol.37 No.2

Background/Aims: Data comparing the antibody responses of different coronavirus disease 2019 (COVID-19) vaccine platforms according to dose with natural severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection-induced antibody responses are limited. Methods: Blood samples from adult patients with mild and severe COVID-19 and healthcare workers who received ChAdOx1 nCoV-19 vaccine (2nd dose at 12-week intervals) and BNT162b2 vaccine (2nd dose at 3-week intervals) were collected and compared by immunoglobulin G immune responses to SARS-CoV-2 specific spike protein using an in-house-developed enzyme-linked immunosorbent assay. Results: A total of 53 patients, including 12 and 41 with mild and severe COVID-19, respectively, were analyzed. In addition, a total of 73 healthcare workers, including 37 who received ChAdOx1 nCoV-19 and 36 who received BNT162b2, were enrolled. Antibody responses after the first and second doses of the ChAdOx1 nCoV-19 vaccine or the first dose of the BNT162b2 vaccine were similar to those in convalescent patients with mild COVID-19, but lower than those in convalescent patients with severe COVID-19, respectively. However, after the second dose of the BNT162b2 vaccine, the antibody response was comparable to that in convalescent patients with severe COVID-19. Conclusions: Our data suggest that the second dose of mRNA vaccination may be more beneficial in terms of long-term immunity and prevention of SARS-CoV-2 variant infection than a single dose of COVID-19 vaccination or homologous second challenge ChAdOx1 nCoV-19.