Generation of a Human Monoclonal Antibody to Cross-Reactive Material 197 (CRM₁₉₇) and Development of a Sandwich ELISA for CRM₁₉₇ Conjugate Vaccines

( Dain Kim ),( Hyeseon Yoon ),( Sangkyu Kim ),( Jimin Wi ),( Heesu Chae ),( Gyunghee Jo ),( Jun-yeol Yoon ),( Heeyoun Kim ),( Chankyu Lee ),( Se-ho Kim ),( Hyo Jeong Hong ) 한국미생물 · 생명공학회 2018 Journal of microbiology and biotechnology Vol.28 No.12

Cross-reactive material 197 (CRM₁₉₇) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. CRM₁₉₇ has been used as a carrier protein for poorly immunogenic polysaccharide antigens to improve immune responses. In this study, to develop a sandwich ELISA that can detect CRM₁₉₇ and CRM₁₉₇ conjugate vaccines, we generated a human anti-CRM₁₉₇ monoclonal antibody (mAb) 3F9 using a phage-displayed human synthetic Fab library and produced mouse anti-CRM197 polyclonal antibody. The affinity (K_D) of 3F9 for CRM₁₉₇ was 3.55 nM, based on Bio-Layer interferometry, and it bound specifically to the B fragment of CRM₁₉₇. The sandwich ELISA was carried out using 3F9 as a capture antibody and the mouse polyclonal antibody as a detection antibody. The detection limit of the sandwich ELISA was <1 ng/ml CRM₁₉₇. In addition, the 3F9 antibody bound to the CRM₁₉₇-polysaccharide conjugates tested in a dose-dependent manner. This ELISA system will be useful for the quantification and characterization of CRM₁₉₇ and CRM₁₉₇ conjugate vaccines. To our knowledge, this study is the first to generate a human monoclonal antibody against CRM197 and to develop a sandwich ELISA for CRM₁₉₇ conjugate vaccines.

Clinical Practice Guideline for Cardiac Rehabilitation in Korea

Chul Kim,성지동,Jong Hwa Lee,Won-Seok Kim,Goo Joo Lee,Sungju Jee,Il-Young Jung,Ueon Woo Rah,Byung Ok Kim,Kyoung Hyo Choi,Bum Sun Kwon,유승돈,Heui Je Bang,Hyung-ik Shin,Yong Wook Kim,Heeyoune Jung,Eung Ju Ki 대한흉부외과학회 2019 Journal of Chest Surgery (J Chest Surg) Vol.52 No.4

Background: Though clinical practice guidelines (CPGs) for cardiac rehabilitation (CR) are an effective and widely used treatment method worldwide, they are as yet not widely accepted in Korea. Given that cardiovascular disease is the second leading cause of death in Korea, it is urgent that CR programs be developed. In 2008, the Government of Korea implemented CR programs at 11 university hospitals as part of its Regional Cardio-Cerebrovascular Center Project, and 3 additional medical facilities will be added in 2019. In addition, owing to the promotion of CR nationwide and the introduction of CR insurance benefits, 40 medical institutions nationwide have begun CR programs even as a growing number of medical institutions are preparing to offer CR. The purpose of this research was to develop evidence-based CPGs to support CR implementation in Korea. Methods: This study is based on an analysis of CPGs elsewhere in the world, an extensive literature search, a systematic analysis of multiple randomized control trials, and a CPG management, development, and assessment committee comprised of 33 authors—primarily rehabilitation specialists, cardiologists, and thoracic surgeons in 21 university hospitals and 2 general hospitals. Twelve consultants, primarily rehabilitation, sports medicine, and preventive medicine specialists, CPG experts, nurses, physical therapists, clinical nutritionists, and library and information experts participated in the research and development of these CPGs. After the draft guidelines were developed, 3 rounds of public hearings were held with staff members from relevant academic societies and stakeholders, after which the guidelines were further reviewed and modified. Results: CR involves a more cost-effective use of healthcare resources relative to that of general treatments, and the exercise component of CR lowers cardiovascular mortality and readmission rates, regardless of the type of coronary heart disease and type and setting of CR. Conclusion: Individualized CR programs should be considered together with various factors, including differences in heart function and lifestyle, and doing so will boost participation and adherence with the CR program, ultimately meeting the final goals of the program, namely reducing the recurrence of myocardial infarction and mortality rates.

Heteronuclear NMR studies on 44 kDa dimer, syndesmos

Kim, Heeyoun,Lee, Inhwan,Han, Jeongmin,Cheong, Hae-kap,Kim, Eunhee,Lee, Weontae Korean Magnetic Resonance Society 2015 Journal of the Korean Magnetic Resonance Society Vol.19 No.2

Syndesmos, which is co-localized with syndecan-4 cytoplasmic domain ($Syn4^{cyto}$) in focal contacts, interacts with various cell adhesion adaptor proteins including $Syn4^{cyto}$ to control cell signaling. Syndesmos consists of 211 amino acids and it exists as a dimer (44kDa) in solution. Recently, we have determined the structure of syndesmos by x-ray crystallography, however, dynamics related to syndecan binding still remain elusive. In this report, we performed NMR experiments to acquire biochemical and structural information of syndesmos. Based on a series of three-dimensional triple resonance experiments on a $^{13}C/^{15}N/^2H$ labeled protein, NMR spectra were obtained with well dispersed and homogeneous NMR data. We present the sequence specific backbone assignment of syndesmos and assigned NMR data with combination structural information can be directly used for the studies on interaction with $Syn4^{cyto}$ and other binding molecules.

High-dose rate γ-irradiation leads to the change of body and organ weights, hematological parameters, frequency of micronuclei in ICR mice

HEEYOUN SHIM,BYULNIM HWANG,YUN-MI BAEK,JEONG-IN KIM,SEO KON KANG,BYOUNG-IL LEE,HEE SUN KIM 한국실험동물학회 2016 한국실험동물학회 학술발표대회 논문집 Vol.2016 No.08

Direct Monitoring of Fucosylated Glycopeptides of Alpha-Fetoprotein in Human Serum for Early Hepatocellular Carcinoma by Liquid Chromatography-Tandem Mass Spectrometry with Immunoprecipitation

Kim, Kwang Hoe,Lee, Soo-Youn,Hwang, Heeyoun,Lee, Ju Yeon,Ji, Eun Sun,An, Hyun Joo,Kim, Jin Young,Yoo, Jong Shin Wiley (John WileySons) 2018 PROTEOMICS CLINICAL APPLICATIONS Vol.12 No.6

Biochemical and NMR Characterization of MTH1880 Mutant Proteins for Folding-Unfolding Studies

Heeyoun Kim,Sooyoung Ryu,윤지혜,김석만,장익수,이원태 대한화학회 2010 Bulletin of the Korean Chemical Society Vol.31 No.12

MTH1880 is a hypothetical protein derived from Methanobacterium thermoautotrophicum, thermophilic methanogen. The solution structure determined by NMR spectroscopy showed that it has a novel α+β-fold with a highly acidic ligand binding pocket. Since MTH1880 maintains its ultra-stable structural characteristics at both high temperature and pressure, it has been considered as an excellent model for studying protein folding. To initiate the structural and folding study of MTH1880 in proving its unusual stability, we performed the site directed mutagenesis and biochemical analysis of MTH1880 mutants. Data from circular dichroism and NMR spectroscopy suggest that the point mutations perturbed the structural stability of protein even though the secondary structure is retained. This study will provide the useful information in understanding the role of participating residues during folding-unfolding process and our result will be used in designing further folding experiments for hyper-thermopile proteins like MTH1880.

Clinical Practice Guideline for Cardiac Rehabilitation in Korea

Chul Kim,성지동,Jong Hwa Lee,김원석,Goo Joo Lee,Sungju Jee,Il-Young Jung,Ueon Woo Rah,Byung Ok Kim,Kyoung Hyo Choi,Bum Sun Kwon,Seung Don Yoo,Heui Je Bang,Hyung-Ik Shin,김용욱,Heeyoune Jung,Eung Ju Kim,Jung Hw 대한재활의학회 2019 Annals of Rehabilitation Medicine Vol.43 No.3

Objective Though clinical practice guidelines (CPGs) for cardiac rehabilitation (CR) are an effective and widely used treatment method worldwide, they are as yet not widely accepted in Korea. Given that cardiovascular disease is the second leading cause of death in Korea, it is urgent that CR programs be developed. In 2008, the Government of Korea implemented CR programs at 11 university hospitals as part of its Regional Cardio-Cerebrovascular Center Project, and three additional medical facilities will be added in 2019. In addition, owing to the promotion of CR nationwide and the introduction of CR insurance benefits, 40 medical institutions nationwide have begun CR programs even as a growing number of medical institutions are preparing to offer CR. The purpose of this research was to develop evidence-based CPGs to support CR implementation in Korea. Methods This study is based on an analysis of CPGs elsewhere in the world, an extensive literature search, a systematic analysis of multiple randomized control trials, and a CPG management, development, and assessment committee comprised of 33 authors—primarily rehabilitation specialists, cardiologists, and thoracic surgeons in 21 university hospitals and two general hospitals. Twelve consultants, primarily rehabilitation, sports medicine, and preventive medicine specialists, CPG experts, nurses, physical therapists, clinical nutritionists, and library and information experts participated in the research and development of these CPGs. After the draft guidelines were developed, three rounds of public hearings were held with staff members from relevant academic societies and stakeholders, after which the guidelines were further reviewed and modified. Principal Conclusions CR involves a more cost-effective use of healthcare resources relative to that of general treatments, and the exercise component of CR lowers cardiovascular mortality and readmission rates, regardless of the type of coronary heart disease and type and setting of CR. Individualized CR programs should be considered together with various factors, including differences in heart function and lifestyle, and doing so will boost participation and adherence with the CR program, ultimately meeting the final goals of the program, namely reducing the recurrence of myocardial infarction and mortality rates.

Identification and quantification by GlycoProteome analyzer (IQ-GPA) in web service

Nahye Kim,Jung Hun Ju,Hyejin Kim,Jin Young Kim,Jong Shin Yoo,Heeyoun Hwang 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07

We have developed an Identification and Quantification by GlycoProteome Analyzer (IQ-GPA) for high throughput analysis of N- and O-glycoproteome. Herein, we introduce the web version of IQ-GPA for a database search and algorithmic suite with tandem mass spectrometry of HCD/CID/ETD (EThcD) in a shotgun experiment. We have designed to easily handle high-throughput glycoproteomic data with graphical user interfaces, and users can instantly access to a range of Glycan Database Builder as well as N and O-GPA modules using Microsoft Azure cloud platforms. With spectra files and the glycopeptide database of interest, the modules identify and quantify intact N- and O-linked glycopeptides in the complex samples, which requires the following steps: 1) DB Builder Module generates a GPA database using the detailed parameters after select working directory. 2) Analysis Module requires a spectrum file and a GPA-Database to analyze N- and O- glycopeptides with user specific parameters. 3) GPA Report Module shows the results and statistical charts on the website. In addition, IQ-GPA contains a module for the classification of N-glycopeptides as core- and outer-fucosylated types using the artificial intelligence ( https://www.iqgpa.org ).

아동학대 사망사건에 대한 신문기사 분석

김지혜(Kim, Ji-Hye),정익중(Chung, Ick-Joong),이희연(Lee, Heeyoun),김경희(Kim, Gyeong-Hui) 한국사회복지학회 2013 한국사회복지학 Vol.65 No.2

학대로 인해 사망하는 아동의 꾸준한 발생에도 불구하고 이에 대한 실태 파악과 대책 마련은 매우 부족한 현실이다. 본 연구는 아동학대 사망사건의 현황과 원인, 특성 등을 확인하기 위해 신문기사에 나타난 아동학대 사망사건을 분석하였다. 연구결과, 2000년도부터 2012년도까지 언론에 보도된 아동 학대 사망사건은 총 141건이었으며, 가해자는 미혼모, 친모, 동거녀 및 계모, 친부의 순인 것으로 나타났다. 아동살해 이유는 미혼모나 미혼부모에 의한 신생아살해의 빈도가 가장 높았고, 다음으로 양육 능력 부족, 산후우울증, 생활고와 경제능력 부족 순인 것으로 나타났다. 또한, 언론에 중점보도된 사건에서 발견된 주제는 범죄현실의 구성-아동학대 사망사건의 가해자에 대한 고정관념, 지속적이고 반복적인 심각한 학대, 신고 및 관리체계 부실, 처벌 미약 등으로 나타났다. 이러한 연구결과를 토대로 본 연구는 아동학대 사망사건을 예방하고 해결하기 위한 제언을 제시하였다. Although fatalities due to child abuse are frequently happened, there has been a dearth of research on the topic. The present research analyzes deaths due to child abuse in representative newspapers in order to examine incidences, causes, and characteristics of child abuse. The results showed that from 2000 to 2012 child abuse deaths occurred 141 cases. Most deaths due to child abuse were committed by single mothers, biological mothers, cohabiting women and stepmothers, and biological fathers. As regards the reasons of deaths due to child abuse, the frequency of newborn infant deaths as the result of abuse by single mother or single parent was the highest, and lack of parenting skills, postpartum depression, hardships of living and lack of financial abilities were followed. Also, subjects received extensive media coverage were crime compositions- stereotypes of child abusers, habitual and continual child abuse, lack of child abuse reporting and prevention system, and mild penalties. Finally, based on the results of the study, the implications to prevent and resolve deaths due to child abuse were discussed.

Site specific characterization of N- and O-Glycosylation in etanercept by TMT-labeling and LC-MS/MS

Hyejin Kim,Heeyoun Hwang,Yeji Jeong,Sangwoo Park,Hyun Joo An,Jin Young Kim,Jong Shin Yoo 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07

Improved methods are required for glyco-characterization of biopharmaceuticals and comparison between their corresponding biologics and biosimilar, where they have multiple glycosylation sites such as Etanercept of a fusion protein. In order to enhance sensitivity of discovery and minimize the sample loss, the combination of a TMT labeling method and fractionation using high flow HPLC has contributed to detect low amount of peptide in data dependent mode of LC-MS/MS. Here, we report a new method combined with TMT labeling, fractionation using ZIC-HILIC HPLC, LC-MS/MS using HCD triggered CID/EThcD, and computational software for identification of site-specific N- and O-glycopeptide and comparison of their corresponding biologics and biosimilar. Of three different Etanercept, such as Enbrel (biologics), Etoloce and Eucept biosimilars), a total of 115 N- and O- glycopeptides were identified and quantified using TMT-11plex labeling including triplicates and 22 ZIC-HILIC fractionation in three and 15 of N- and O- glycosylation sites, respectively. Two O-glycopeptides in different O-glycosylation site and seven core 2 type of O-glycopeptides were first reported. In particular, three kinds of O-glycopeptides with NeuGC which could be a foreign agent of immune response. In depth of quantification, our strategy shows site-specific similarity is quite not similar between biologics and biosimilar.