Commercial-scale Economic Comparison of Different Batch Modes for Upstream and Downstream Processing of Monoclonal Antibody

차현명,Hwang Jeong-Min,연정흠,임진혁,한혜진,김동일 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.6

Chinese hamster ovary (CHO) cell cultures are widely used due to their high productivity, industrial track record, safety record, and correct post-translational modifications. Fed-batch and perfusion modes have been mostly adopted as traditional commercial manufacturing process using CHO cells for the production of biopharmaceuticals. Thus, we compared fed-batch and perfusion culture processes for the commercial-scale production of monoclonal antibody. Main production process was performed after confirming the comparable culture performance in seed train of different operation modes, and then perfusion cultures showed 1.6- fold higher peak viable cell density (VCD) with a longer culture duration than that of fed-batch cultures. Also, as the result of main batches, total product amount increased over 450% in perfusion cultures. Importantly, perfusion process allowed to improve the higher purification yield while maintaining acceptable product quality, because the level of process impurity could be reduced by using retention device. When economic comparison of fed-batch versus perfusion processes was carried out under the equal scale of upstream and downstream, perfusion mode was more a cost-effective bioprocess that could reduce the cost of goods. In conclusion, although comparison data of triplicate commercial batches are of course specific case for a given antibody production, this study verified the possibility of using perfusion mode as an efficient system for commercial producing the desired antibody and is a meaningful report in the biopharmaceutical industry.

중간엽줄기세포 증식을 위한 누에 실샘 유래 무혈청 배지 첨가제

차현명 ( Hyun Myoung Cha ),김선미 ( Sun Mi Kim ),최용수 ( Yong Soo Choi ),박지성 ( Ji Sung Park ),임진혁 ( Jin Hyeok Lim ),황슬기 ( Seul Gee Hwang ),김동일 ( Dong Il Kim ) 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.1s

Mesenchymal stem cells (MSCs) are must be cultured in chemically defined, xeno-free culture system for cell-based therapies. One major overcome for their clinical use is the unsafety of fetal bovine serum (FBS), which is a crucial part of all media currently used for the culture of MSCs. Previously, we developed novel methods the production of silkworm gland hydrolysate (SGH). Also, we showed the effect of the SGH on the cell growth promotion. We demonstrated whether the characteristics of MSCs were maintained in the serum-free media containing SGH during long-term cultivation. The results of microscopic observation showed that the morphology of stem cells were not different between the media containing 10% FBS and 1 mg/mL SGH. After the long-term culture of MSCs in serum-free media containing SGH, the rate of MSCs growth was well maintained. The analysis of anti-apoptotic effect of SGH showed that early apoptosis of MSCs was reduced by 3.7-fold. Furthermore, MSCs specific surface markers and stemness gene expression (Nanog, Sox2) were preserved as that of MSCs in the media containing 10% FBS. In conclusion, we demonstrate that the SGH can be used as useful supplement for MSCs cultivation in serumfree media.

한국판 애도 관련 회피 척도(K-GRAQ)의 타당화

차성이,현명호 한국건강심리학회 2024 한국심리학회지 건강 Vol.29 No.3

본 연구는 사별 경험 이후 죽음과 상실을 상기시키는 자극에 대한 회피를 측정하는 애도 관련 회피 척도(GRAQ)를 국내 실정에 맞게 번안하고 타당화하기 위해 진행하였다. 이를 위해 원 저자의 동의와 검토를 거쳐 한국판 애도 관련 회피 척도(K-GRAQ)의 최종 문항을 구성하였다. 이후 친밀한 대상과의 사별을 경험하고 6개월 이상 20년이 경과하지 않은 만 19세 이상 60세 미만의 성인 483명을 대상으로 척도의 신뢰도와 타당도를 검증하였다. 척도의 내적 일치도와 문항 총점 상관, 검사 재검사 신뢰도를 검토한 결과 K-GRAQ의 신뢰도는 양호한 것으로 나타났다. K-GRAQ의 요인 구조를 확인하기 위해 탐색적 요인분석(n=241)과 확인적 요인분석(n=242)를 실시하였다. 탐색적 요인분석 결과 2개 요인이 도출되었으나, 2요인 모형의 적합도보다 원저자가 제안한 3요인 모형의 적합도가 더 양호한 것으로 확인되었다. 또한, K-GRAQ와 복합 비애 증상과 우울, 불안 증상과의 상관분석을 통해 수렴 타당도를 검증하였다. 마지막으로 본 연구의 의의와 한계점을 제언하였다. This study aimed to validate the Grief-Related Avoidance Questionnaire(GRAQ) developed by Shear et al(2007) according to domestic situations. The final items for the K-GRAQ were constructed with consent and review from the original authors. Subsequently, the reliability and validity of the scale were tested on 483 adults aged between 19 and 60 who had experienced bereavement in intimate relationships between 6 months and 20 years of the loss. The reliability of the K-GRAQ was deemed satisfactory, as evidenced by its internal consistency, total-item correlations, and test-retest reliability. Exploratory factor analysis (n=241) and confirmatory factor analysis (n=242) were conducted to confirm the factor structure of the K-GRAQ. While exploratory factor analysis revealed a two-factor solution, the fit indices favored the three-factor model proposed by the original authors. Additionally, convergent validity was confirmed through correlation analysis between the K-GRAQ and the symptoms of complicated grief, depression, and anxiety. Finally, the significance and limitations of this study were discussed.

복합 비애(Complicated Grief)에 대한 국내 연구동향(2010~2020): 체계적 문헌 고찰

차성이,박신영,현명호 한국심리학회 2022 한국심리학회지 일반 Vol.41 No.4

This study aimed to review the literature on Complicated Grief(CG) in South Korea for 10 years from 2010 and summarized the significant results. 33 papers out of 188 studies were selected through a systematic literature review. As a result, the majority of the studies conducted after 2015, and most were classified as quantitative studies. When looking at the characteristics of the study, 19 studies considered the elapsed period after bereavement, 28 studies identified the subject of bereavement. Inventory of Complicated Grief(ICG) and Prolonged Grief Disorder-13(PG-13) were most popular measures for CG but the use of validated measurement were limited. Factors related to CG were included attachment type, intrusive rumination, and perceived social support. Protected factors affecting the coping process of bereavement were future-oriented coping style, coping flexibility, and meaning-making process. Additionally, we summarise the results of the study on traumatic loss and the studies examine the effectiveness of the psychological intervention program on the recovery of CG. Lastly, we discussed the limitations of the literatures and the potential implications and suggestions for further research. This study is significant in that it is the first study to systematically examine studies conducted on CG in South Korea. 본 연구는 2010년부터 10년간 국내에서 한국인 성인을 대상으로 이루어진 복합 비애(Complicated Grief)에 대한 연구문헌을 분석하여 결과를 요약하고 연구 성과와 한계점을 살펴보고자 하였다. 이를 위해 최근 10년간 복합 비애에 대해 연구한 188편의 논문을 검색하고 체계적 문헌 고찰 방법을 통해 최종 33편을 대상으로 선정하여 분석하였다. 그 결과, 연구 대상 문헌 중 2015년도 이후에 진행된 연구가 28편으로 대다수였으며 대부분 양적 연구로 분류되었다. 또한, 사별 후 경과 기간을 고려한 연구는 19편, 사별 대상을 파악한 연구는 28편, 사별 유형을 파악한 연구는 22편이었다. 측정 도구는 복합 비애 척도(ICG)와 지속성 비애 장애 척도(PG-13)가 주로 사용되었으나 타당화된 척도를 사용한 연구는 부족했다. 또한, 복합 비애에 영향을 미치는 요인으로는 애착 유형, 침습적 반추, 지각된 사회적 지지가 있었으며, 복합 비애의 대처 과정에 영향을 미치는 요인으로 미래 중심적 대처와 대처 유연성, 의미 재구성이 있었다. 추가적으로 외상성 상실과 복합 비애에 대한 연구 결과와 복합 비애의 회복에 대한 심리 개입 프로그램의 효과성을 탐색한 연구를 요약하였다. 마지막으로 연구 대상 문헌의 한계점과 향후 복합 비애에 대한 보다 폭넓은 연구 필요성에 대해 제언하였다. 본 연구는 국내에서 복합 비애에 대해 진행한 연구를 체계적으로 고찰하여 연구동향을 파악한 최초의 연구라는 점에서 의의가 있다.

Mediating Effects of Attentional Control in the Relationship Between Neuroticism and Repetitive Negative Thinking

차은정,현명호 한국임상심리학회 2021 Korean Journal of Clinical Psychology Vol.40 No.2

Depression and anxiety symptoms frequently co-occur. Research suggests that a common shared symptom — repetitive negative thinking (RNT) — is a vulnerability factor that develops and prolongs symptoms of affective disorders. A dispositional factor that increases RNT is neuroticism. The mental noise hypothesis of neuroticism suggests that its volatile and reactive nature causes attentional control deficits. As theories of RNT, specifically regarding rumination and worry, indicate such deficits as underlying causes, it is hypothesized that the deficits may mediate the relationship between neuroticism and RNT. This study investigated whether attentional focusing and shifting mediates the relationship between neuroticism and RNT (worry and rumination). Results showed mediational effects of focusing on rumination, while the results on worry were not significant. Neuroticism has been discussed as a temperamental risk factor that increases vulnerability to psychopathology. Future research should employ longitudinal designs and behavioral measures to overcome this study’s limitations.

군대 내 대인 외상 피해자를 위한 트라우마 단기 상담 프로그램 예비 연구

박신영,차성이,현명호 대한스트레스학회 2022 스트레스硏究 Vol.30 No.3

Background: This preliminary study was conducted to develop a short-term counseling program for victims who experienced interpersonal trauma in the military and to verify its effectiveness. Methods: A single group pre-post-mixed design was used for this study. Seven victims who experienced interpersonal trauma within the military participated in the program once a week for five weeks. Participants’ post-traumatic stress symptoms, degree of insomnia, depression, state anxiety, and acceptance behavior levels were assessed and compared at baseline and pre-and post-test. Results: Participants’ post-traumatic stress symptoms, state anxiety, and depression symptoms significantly reduced and acceptance behavior significantly increased after participating in the short-term counseling program. Conclusions: The results of this study will help explore therapeutic elements that are helpful for victims who have experienced interpersonal trauma in the military. 본 연구는 군대에서 대인 외상을 경험한 피해자를 대상으로 트라우마 단기 상담 프로그램을 개발하고자진행한 예비 연구이다. 본 연구에서는 트라우마의 단계 기반 치료적 접근 중 1단계에 기초한 프로그램을구성하여, 참여자(n=7)를 대상으로 매주 1회 진행하였다. 참여자의 외상 후 스트레스 증상, 불면증 정도, 우울, 상태 불안 및 수용 행동 수준을 기저선, 사전 및 사후와 추수 시점에 측정하여 비교한 결과, 외상후 스트레스 증상, 상태 불안, 우울 증상이 감소하였고 수용 행동은 증가하였다. 본 연구 결과는 군대에서대인 외상을 경험한 피해자를 위한 치료 요소를 탐색하는 데 도움이 될 것이다

CHO 세포 배양에서 Hexose 혼합 비율 조절에 의한 Albumin-erythropoietin의 Sialylation 증대

김나희,차현명,임진혁,이지훈,한혜진,김동일 한국생물공학회 2019 KSBB Journal Vol.34 No.2

Sialylation is very important for in vivo half-life oftherapeutic proteins because sialic acids can reduce the ratesof clearance in hepatocyte. Many studies were developed forsupplementing various carbon sources which directly influcencedcellular metabolism and glycosylation. So it is necessaryto investigate interconversion of carbons sources in combinationbecause once they are absorbed into cells and synthesizedinto nucleotide sugars, they can be converted to differentkinds of sugars and nucleotide sugars. Albumin-erythropoietin(Alb-EPO) has three glycan sites with tetra-antennarystructure, so it can be added up to 14 sialic acids. Therefore,we demonstrated the effects of combination ratios of hexose(glucose:galactose:mannose) among the generally used carbonsources essential for cell growth on sialylation of a fusionprotein, Alb-EPO. As a result, there was no negative effect oncell growth except the condition of the highest concentration. The titer of Alb-EPO was increased depending on increasingconcentration of hexoses in order of mannose, glucose, andgalactose and reached up to 1.7-fold in comparison to the control. On the other hand, there was more than 1.5-fold increasein sialylation followed by changed nucleotide sugar pool atthe conditions supplemented with glucose and galactose athigh concentration. In conclusion, our strategy of controllingthe concentrations of available carbon sources offers an effectiveapproach for the improved production of highly sialylatedAlb-EPO without deleterious effect on growth.

아동기 어머니의 정서적 학대와 복합 외상후 스트레스 증상 및 용서의 관계

라영선,현명호,차성이,윤선영 한국임상심리학회 2010 Korean Journal of Clinical Psychology Vol.29 No.1

본 연구는 아동학대 중에서도 어머니의 정서적 학대가 복합 외상후 스트레스 증상을 예측하는지와 정서적 학대와 복합 외상후 스트레스 증상의 관계에서 용서의 조절효과에 대해 알아보고자 하였다. 용서는 특질용서와 상황용서로 구분하여 각각의 효과를 살펴보았다. 이를 위해 대학생을 대상으로 정서적 학대, 특질용서, 상황용서, 그리고 복합 외상후 스트레스 장애 설문지를 실시하였다. 그 결과, 정서적 학대는 복합 외상후 스트레스 증상을 예측하는 것으로 나타났다. 위계적 회귀분석 결과, 정서적 학대와 복합 외상후 스트레스 증상의 관계를 상황용서와 특질용서가 조절하였고 특질용서보다 상황용서의 효과가 더 크게 나타났다. 본 연구 결과는 아동기에 정서적 학대를 경험한 정도가 높은 사람일수록 복합 외상후 스트레스 증상이 심각하다는 것을 나타낸다. 또한 정서적 학대가 심각하더라도 상황용서와 특질용서 수준이 높으면 복합 외상후 스트레스 증상이 완화될 수 있다는 것을 시사한다. 마지막으로 연구의 의의와 제한점 및 추후연구에 대한 방향을 제시하였다.

Utilization of Glucocorticoids as Additives for Enhanced Sialylation of Fc-fusion Protein in CHO Cell Cultures

한혜진,임진혁,차현명,이지훈,김동일 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.2

Sialylation, which is considered as a critical quality attribute, plays important roles in pharmacokinetics of recombinant proteins. Terminal sialic acids of glycoproteins are modulated by intracellular sialyltransferase (ST) and extracellular sialidase. Since CHO cells producing recombinant proteins can only transfer α2,3-sialic acid to a nascent oligosaccharide, inactivation of α2,6-ST results in a different composition of glycan with human proteins, which have predominantly α2,6-linkages. To overcome this problem and maximize total sialic acid contents, we used glucocorticoids (GCs) that are classified as a steroid hormone in CHO cells expressing both α2,3-ST and α2,6-ST. This study was performed to determine the effect of GCs on CHO cell cultures and sialylation of Fc-fusion protein. We observed that all cultures supplemented with GCs reduced the titer of the Fc-fusion protein, but enhanced the sialylation level, when compared to those of control. Especially, addition of corticosterone increases the levels of α2,6-sialylation as well as the total contents of sialic acid. Enhanced sialic acid contents are thought to be due to the intracellular improvement by sialylatransferase and the inhibition of sialidase. In conclusion, GCs can be utilized as an effective medium additive to improve the quality of biopharmaceuticals using CHO cell cultures.

CHO 세포 배양을 통한 Recombinant Human Erythropoietin의 생산에서 저혈청 배지와 배양 첨가물질이 미치는 영향

이경선(Kyung-Sun Lee),차현명(Hyun-Myoung Cha),임진혁(Jin-Hyuk Lim),김동일(Dong-Il Kim) 한국생물공학회 2017 KSBB Journal Vol.32 No.2

Mammalian cell cultures have been used extensively to produce proteins for therapeutic agent because of their ability to perform post-translational modification including glycosylation. To produce recombinant protein, many factors and parameter are considered such as media composition, host cell type, and culture process. In this study, recombinant human erythropoietin (rhEPO) producing cell line was established by using glutamine synthetase system. To reduce serum concentration in media, we compared direct adaptation with step adaptation. Cell growth was faster in step adaptation. In low-level serum media, there were insufficient glucose for cell growth. Thus, we added glucose in low-level serum media from 2 g/L to 4.5 g/L. Titer of rhEPO was higher than other conditions at 4.5 g/L of glucose. Additionally, Nmethyl-D-aspartate (NMDA), 13-cis-retinal, and pluronic F-68 (PF-68) were added to enhance productivity in CHO cell cultures. In conclusion, we applied CHO cell producing rhEPO to low-level of serum in media using step-adaptation. Also, we confirmed positive effect of NMDA, 13-cis-retinal, and PF-68.