Development of Nivolumab Bio-Better using mammalian and plant Glyco-Delete platform

강초은 Graduate School, Yonsei University 2022 국내석사

항체에 부착된 당사슬은 다양한 생물학적 활성, 즉 단백 안전성, 이동성, 단백 제거율, 수용체 접합성 등을 변화시켜 치료 효능, 체내 지속성, 타겟팅 및 면역반응 등에 중요한 역할을 수행하기 때문에 항체의약품의 품질을 결정하는 주요인자이다. 그러나 당사슬은 다양한 효소의 작용으로 만들어지고 생산공정 과정이나 조건에 따라 각 효소의 활성이 다를 수 있으므로 단백질에 균일하지 않게 부착될 수 있다. 이러한 당사슬을 균일하게 조절할 수 있다면 항체의 품질을 지속적으로 유지할 수 있다. 특히 표적가림용 항체의약품의 경우 항체부착에 의해 표적발현 세포의 사멸을 초래할 수 있는데, 당사슬 조절을 통해 그러한 표적발현세포의 사멸 능력을 제거할 수 있다. 본 연구는 CHO 세포와 식물인 담배의 당화과정을 조절하여, 항체의 ASN297에 한 개의 GlcNAc 분자만 부착된 형태를 생산하는 당사슬제거 (Glyco-Delete, GD) CHO세포와 당사슬제거 담배시스템을 구축하고 이를 통해 생산된 항체의 기능을 평가하는 연구이다. IgG1과 IgG4 형태의 니볼루맵들을 생산하여, 고유능력인 항원결합력, PD1 활성화 예방능력 및 싸이토카인 분비 등의 T 세포 활성조절능력을 확인할 뿐 아니라, 궁극적으로 PD1발현 세포에 대한 면역원성의 감소를 관찰하는 것을 목적으로 했다. 또한 당사슬제거 식물 시스템을 이용하여 마찬가지로 식물 니볼루맵을 생산하여 그 기능을 함께 평가하고자 했다. 본 연구는 CHO 세포 및 식물에서 생산된 당사슬제거 니볼루맵들의 면역원성이 낮아짐을 증명했다. 이를 통해 니볼루맵의 타겟인 T세포의 사멸을 줄여 개선된 안정성을 규명함과 동시에, 항체 적용 시 면역활동에 참여하여 효과를 일으키는 T세포의 사멸을 줄임으로써 궁극적으로 향상된 항체의 효능 또한 규명하였다. 이러한 결과는 니볼루맵과 같은 표적가림용 항체의 성능을 향상시켜 동∙식물 항체 플랫폼의 개선에 기여할 수 있을 것이다. The glycan in Fc region regulates therapeutic efficacy of antibody by changing various biological activities such as protein safety, mobility, protein clearance, and receptor conjugation. Also, since it has various biological activities, such as playing an important role in targeting antigen and immune response, it is a factor that determines the quality of antibody. However, since glycans are made by the action of various enzymes and the activity of each enzyme may be different depending on the production process or conditions, it is difficult to homogeneously attach to the protein. Nivolumab is a human IgG4 monoclonal antibody that blocks PD1. It was approved for medical use in the United States in 2014 and treats several types of cancer includes melanoma, lung cancer, malignant pleural mesothelioma, renal cell carcinoma, etc. It is mainly used as a first-line treatment for metastatic melanoma and non-small cell lung cancer, and many clinical trials are ongoing as a combination treatment with various targeted anticancer drugs. Also, Nivolumab is the type of immunotherapy works as a checkpoint inhibitor, blocking a signal that prevents activation of T cells from attacking the cancer. Most immunomodulatory anticancer drugs including nivolumab have IgG4 backbone which has low affinity for complement and various FcγRs. However, IgG4 still has a measurable affinity for FcγRIII, so given the high drug concentrations used in therapy, the possibility of target cell ADCC cannot be completely eliminated. Therefore, a method for removing immunogenicity of the IgG4 is highly demanded. Glyco-Delete (GD) technology is an ADCC removal platform for antibody drugs. Via glycosylation enzymes regulation to leave only one GlcNAc glycan stump, not only reduce the affinity for FcγRIII, but also provide the uniformity of antibody which is important factor for manufacturing. In this study, GD CHO cells and GD plant systems were produced by controlling the glycosylation process of CHO cells and plant and nivolumab was used for template to prove the usefulness of GD platform. By producing nivolumab with IgG1 and IgG4 backbone in CHO cell system, we measured the affinity for PD1 and the blocking activity of PD1 signaling to check whether the GD cause any change in affinity and blocking activity of PD1 signaling. Glycan uniformity and immunogenicity were also confirmed the improved characteristics of GD nivolumab. Furthermore, by using the GD-plant system, plant-nivolumab was produced and its functions were evaluated. This study demonstrates that GD platform of CHO cells and plant provides reduced ADCC activity to nivolumab without altering PD1inhibiting activity. GD engineering can be contributed to as a bio-better platform for various target-masking antibodies.

Characterization of the structural and functional aspects of the animal lectin, galectin-1

조문제 University of Oklahoma 1995 해외박사

Chinese Hemster Ovary(CHO) 세포로 부터 Galectin-1 cDNA를 만들어 이를 대장균에 이식 recombinant galectin-1 (rGal-1)을 대량 생산하였고 두번째 Cysteine이 Serine으로 변형된 C₂S Gal-1도 같은 방법으로 생산하였다. 환원제가 첨가된 용액에서만 그 활성을 유지하는 것으로 알려진 Gal-1은 두번 째 Cysteine이 Serine으로 변화된 경우 더이상 수용액내의 환원력에 의해 그 활성이 지배받지 않았다. 한편 rGal-1은 그들의 결합상대인 Polylactosamine 구조의 Oligo당과 함께 수용액상에 저장하는 경우 수용액내의 환원력이 또한 생화학적 활성에 영향을 주지 못하였다. 예상치 못한 Gal-1은 monomer형태와 dimer형태로 존재하며 이는 농도에 따라 결정되는 가역적 평형을 이루고 있었 다 (kd=-7uM). 이 두가지 형태는 모두 생화학적 활성을 가지고 있으며 이러 한 현상은 rGal-1 뿐만 아니라 CHO세포에서 직접 분리한 native Gal-1에서도 사실임이 판명되었다. Gal-1의 생체내 기능이 그들의 dimer형성에 의한 multivalancy에 의존하는 경향이 있으므로 농도에 관계없이 monomer로 존재할 수 있는 monomeric mutan t를 만들게 되었다. Gal-1의 dimer형성은 X-ray Crystallography를 분석한 결과 각 분자의 N-terminal과 C-terminal에 접촉에 의한 비공유성 결합인것 으로 사료되어, N-terminal의 3개의 친유성 아미노산을 친수성 아미노산으로 대체하였다. 그 결과 고농도에서도 dimer가 형성되지 않는 monomeric Gal-1 을 만드는데 성공하였다. 세포내에서 생성된 Gal-1의 존재위치는 반은 세포 표면이며 이는 세포막 표면에 존재하는 당단배질의 당 구조와 결합하는 형태 로 존재하는 것을 알 수 있었다. 위와 같은 결과는 물리적 특성조사에서 밝 혀진 사실과 서로 보완되는 것으로 세포 외 표면에서는 세포내부에 비하여 환 원력이 떨어진 상태로 Gal-1이 당 구조와 결합하지 않는 경우 쉽게 그 활성이 상실된다는 것이다. 이는 또한 Gal-1이 결합하는 당 구조를 만들지 못하는 변이주(Lec 8CHO)세포를 이용한 실험에서 Gal-1이 세포 표면에서 발견되지 않 은 사실로도 증명되었다. Galectins are the soluble, O-galactoside binding lectin found in various kinds of tissues in vertebrate and invertebrate. The function of galectin is still unclear partially because protein itself is not well characterized expect its binding property. In order to study the relationship between structure and proposed function, I choose galectin-1 and CHO cells for model system because galectin-1 is the prototype of galectin family and CHO cells produce relatively large amuont of galectin-1. cDNA for galectin-1 from CHO cells was prepared and sequenced and a recombinant form (rGal-I) was expressed in E. co/i. A mutated form of the protein that fully retained activity was also constructed (termed C2SrGal-1) in which Cys-2 was changed to Ser-2. rGal-I was stable in the presence of reducing agent, but it quickly lost all activity in the absence of reducing agent. In contrast glycoprotein ligands, such as basement membrane laminin, stabilized the activity of rGal-1 in the absence of reducing agent (tt/s >2 weeks). C2SrGal-1 was stable in the presence or absence of either ligand or reducing agent. Unexpectedly, galectin-1 was found to exist in a reversible and active monomer-dimer equilibrium with a Kd=7 PM and an equilibration time of tt/s=lO h. Addition of haptenic sugars did not affect this equilibrium. Galectin-1 isolated from the cytosol of CHO cells was found to exist as monomers and dimers. These studies demonstrate that galectin-1 binding to a biological ligand stabilizes its activity and that the monomer/dimer state of the protein is regulated by lectin concentration. The other controversal aspects of galectin-1 is its localization. It has been known to be synthsized and stay in cytosol. To function as lectin, it has to be secreted to bind to carbohydrate, but its extracellular existence has been controversal. I have now used several approaches to define the subcellular location and biosynthesis of galectin-1 in these cells. Galectin-1 was present on the ceil surface, as assessed by immunofluorescent staining with monospecific antibody to the protein. Quantitation of the surface-localized galectin-1 was achieved by metabolically-radiolabeling cells with saS-MeffCys and measuring the amount of lectin (i) sensitive to trypsin, (ii) accessible to biotinylating reagents, and (iii) accessible to the haptenic disaccharide lactose. By all three procedures =1/2 of the radiolabeled galectin-I associated with cells was shown to be on the cell surface with the remainder intracellular. The kinetics of externalization of galectin-1 was monitored by pulse/chase radiolabeling and it was shown that cells secrete the protein with a tt/s=2Oh. The cell surface form of galectin-1 in CHO cells was active and bound to surface glycoconjugates, but lectin accumulating in the culture media was inactive. Lectin synthesized by mutant Lec8 CHO,cells, which are unable to galactosylate glycoproteins, was not found on the surface and quantitatively accumulated in the media in an inactive form. Taken together, our results demonstrate that galectin-1 is quantitatively externalized by CHO cells and can associate with surface glycoconjugates where the lectin activity is stabilized. The functions of galectin-1 are not yet completely understood, but it is presumed that dimerization and multivalency are important. The crystal structure of bovine galectin-1 predicts N- and C-terminal interactions at the subunit interface. Here we report the construction of galectin-t mutated in residues at the subunit interface. One of these mutants (N-Gal-l) contains disruptions of 3 hydrophobic amino acids at the N-terminus. A second mutant (N/C-Gal-i) was constructed, in which hydrophobic amino acids in both the N-and C-termini were altered. N-Gal-l is monomeric at concentrations up to 80 PM, whereas the native form of the lectin is dimeric at this concentration. Most importantly, N-Gal-l is a functional l3-galactoside-binding protein and binds avidly to laminin. N/C-Gal-l is also monomeric, but it lacks detectable carbohydrate binding activity. These results demonstrate that N-terminal residues are critical for dimer formation, but do not directly contribute to carbohydrate binding activity. The availability of active and inactive monomeric forms of galectin-I may aid in future studies aimed at understanding the biological functions of the lectin and the role of multivalency.

Memory on a platter : food, family, and loss in Julia Cho's plays

Yang, Na-young Graduate School, Yonsei University 2018 국내석사

음식은 인간의 생명과 직결되는 생존을 위한 도구임과 동시에 다양한 함의를 지니는 상징이기도 하다. 음식을 먹는 것 혹은 먹지 않는 것은 종종 인간의 감정을 드러내는 매개체가 된다. 뿐만 아니라 배고픔은 정서적 결핍을 나타내는 은유로 사용되어 왔다. 본 논문은 줄리아 조의 작품 속에서 나타나는 '음식'을 중심으로 가족, 정체성, 상실과 트라우마를 살펴보고자 한다. 무엇을 먹거나 마시는지, 혹은 먹지 않거나 마시지 않는지를 통해 각 인물들의 정체성과 감정을 살펴보고, 그것이 어떻게 물리적 공간과 시간을 벗어난 개인 혹은 가족의 치유와 화해로 이어지는지 살펴본다. 가족 구성원들은 음식 섭취와 요리를 통해 기억과 감정을 공유하고 서로를 이해하며 세대 간의 갈등을 극복한다. 음식을 통해 과거와 현재는 연결되고, 망자는 기억되며, 아픔은 천천히 소화된다. 또한 <The Architecture of Loss>, <Durango>, <Aubergine>에 공통적으로 등장하고 있는 부엌과 그 이외의 공간을 비교하여 음식이 가지는 공간성에 대해 연구하고, 그것이 인물들에게 어떤 시간적 의미와 영향을 가지는지 살펴본다. 더 나아가, 가족의 결핍, 어머니의 부재와 대체, 시공간을 벗어난 집(home)의 의미에 대해 연구하였다. 아시아계 미국인의 인종 정체성을 부정하지 않으면서도 그것에 집착함으로써 개인의 복합적 정체성과 삶의 기억을 인종으로만 한정 짓지 않는다는 점에서 줄리아 조의 작품들은 현대 아시아계 미국 희곡이 직면한 딜레마이자 앞으로의 행보를 접시에 담아낸다. Anyone would have experienced the moment when a whiff or a taste of food instantly transports him/her to the past that has been preserved in his/her memories. This project starts from questioning the relationship between food and memory which is the sine qua non of shaping who I am and whom I am with in Julia Cho’s plays. While Cho portrays precarious family histories and relations, food emerges as a significant symbol imbued with memories and identity that deftly transcends the in and out, visible and invisible, and tangible and intangible. As a thematic and textual study, this thesis examines how Cho's plays—mainly but not restricted to The Architecture of Loss, Durango, and Aubergine—suggest a reconciliation of generations, of individuals, and of life and death, through culinary experiences and palatal memories. Food fosters and connects the relationships between the protagonists while demonstrating their ontological struggle with uneasy reality, bereavement, and familial conflict. Eating becomes a personal history; cooking becomes a symbolic gesture of affection, apologies, and compassion—all of which words often fail to fully convey. The goal of this project is to grapple with the elusive yet, ever-present role that food plays in one’s memory of family and of home.

A method of Efficient Prediction Analysis of Sleep Duration in Smart Home Based on Using PIR sensors

CHO HILARY SCOTT NKIMBENG 아주대학교 아주대학교 일반대학원 2018 국내석사

Activities of Daily Living(ADL) refer to the activities carried out by individuals in their everyday living. ADL’s are good indicators of the health status of individuals. Proper monitoring of these activities can be achieved by attaching state-change sensors to objects in the home which gives a reflection of the object interaction, usage and subsequently the ongoing activity. The increase in the elderly Population nowadays will lead to an increase in the cost of elderly care.Also there is increase decline in sleep durations nowadays short sleep durations (≤6h) could have Cardiovascular disease risk than long sleep durations(≥7h).In this thesis by using PIR sensors and other Binary sensors from Uci binary data set and our proposed fuzzy logic model expert system we efficiently predicted activities of daily living and Sleeping duration in the home based on the acquired binary sensor data. We obtained 96.5% accuracy for Daily Living Activity Prediction and 100% accuracy Sleep duration based on our Fuzzy logic model and binary sensors data. Conclusively, with our method we can effectively predict daily living activity in the home and the sleeping duration class of the elderly home user as a means of improving the care situation of the user.

Imprint- and plasma-etch-patterned organic light-emitting diodes with through-hole anode connections

Cho, Jaewan Sungkyunkwan university 2020 국내석사

포토리소그래피 없이 마이크로미터 크기의 유기 발광 다이오드 (OLED) 픽셀이 제작되었다. OLED 픽셀은 임프린트 리소그래피 및 플라즈마 식각을 사용하여 40 μm 크기로 패터닝되었다. 임프린트 스탬프에 의해 압착된 임프린트 레진은 3차원 4층 구조를 가지며 각 층을 순서대로 식각하여 OLED 픽셀이 제작되었다. 제작된 OLED 픽셀을 별도로 구동하기 위해, 각 픽셀에 스루홀 (through-hole)을 뚫고 금속을 채워 양극 전극이 음극 전극과 동일한 평면에 위치하도록 하였다. 양극과 음극 전극은 금속 lift-off 공정을 통해 분리되었다. 공정의 타당성은 동일한 공정을 통해 동시에 제작된 더 큰 OLED 픽셀을 구동시키는 방법으로 검증되었다. 본 연구에 제시된 방식은 대면적 OLED 디스플레이 제조에 적용될 수 있다. A micron-sized organic light-emitting diode (OLED) pixel array was fabricated without the use of photolithography. The OLED pixels were patterned down to a size of 40 μm using imprint lithography and plasma etching. Molded imprint resist pressed by imprint stamp has a three-dimensional, four-story structure and the OLED pixels were fabricated by removing each layer in order. In order to separately drive OLED pixels, a through-hole was drilled in the pixel and filled with an anode metal so that the anode electrode was located on the same plane as the cathode electrode. The anode and cathode electrodes were separated by a simple metal lift-off process. The feasibility of the fabrication process was verified by operating a larger OLED pixel fabricated simultaneously by the same process. The simple method proposed in this study could be applied to manufacturing large area OLED display.

Stability enhancement of solution-processed oxide thin film transistors based on rare-metal free oxide materials and channel structure design

Cho, Sungwoon Sungkyunkwan university 2018 국내박사

For practical solution-processed oxide thin-film transistor (TFT) with cost efficiency, high performance, and long-term environmental reliability, we suggested novel sol-gel processed rare-metal-free oxide-based semiconductor/insulator [ZnSnO (ZTO)/Al2O3] heterostructure channel and chemically stable sol-gel multi-stacking method. In the rare-metal-free ZTO/Al2O3 heterostructure, the In- and Ga-free ZTO semiconductor with high chemical durability is employed as an effective electron transport layer. Earth-abundant Al2O3 insulator is employed as both an ambient gas molecule barrier at the ZTO back-channel region and a tunneling-induced electron transport layer beneath the source/drain electrodes. In order to minimize inevitable chemical attack coming from acidic and basic precursor solutions during the sol-gel based heterostructure construction, chemically stable sol-gel ZTO/Al2O3 heterostructure stacking was successfully demonstrated with a chemically durable Sn-modulated ZTO semiconductor and weakly corrosive pH-engineered Al2O3 precursor solution. The proposed rare-metal-free ZTO/Al2O3 heterostructure and chemically stable stacking realized sol-gel processed oxide TFT with excellent stability under humidity, temperature, bias voltage, and light exposure. We believe that our novel ZTO/Al2O3 heterostructure and chemically stable sol-gel stacking method will provide an interesting route for the fabrication of practical solution-based oxide TFTs with cost efficiency, high performance, and long-term reliability, instead of conventional rare-metal based oxide materials and channel structures with high environmental instability. In chapter 2, we confirmed that it is possible to minimize the defects in the solution-coated oxide semiconductor and improve the chemical resistance by controlling the heat treatment temperature and time and tin composition of the solution process oxide semiconductor. This resulted in the fabrication of high stability / low cost zinc tin oxide semiconductors, which enabled stable solution stacking without corrosion during oxide semiconductor / insulating layer based heterostructure stacking In chapter 3, we have investigated the corrosion behavior of oxide semiconductor thin films and the corrosiveness of solution precursors for the chemically stable solution-based multilayer laminating process design of oxide semiconductors/insulator-based heterostructures. The changes in corrosion properties and chemical durability of zinc tin oxide of high-endoxicity in pH-controlled solutions were experimentally evaluated and confirmed to be theoretically interpretable and predictable based on the pourbaix-diagram. In chapter 4, We have fabricated a new concept solution-based oxide semiconductor / insulator heterostructure-based high-stability/low-cost transistor by realizing a non-rare-metal oxide-based oxide semiconductor/insulator-based heterostructure through a chemically stable solution construction. This complete solution-based oxide transistor device showed high stability over environmental stresses (moisture, temperature, light, repetitive voltage).

Sidewall patterning of organic-inorganic multilayer thin film encapsulation by adhesion lithography

Cho, Hyejin Sungkyunkwan university 2021 국내석사

For highly flexible Organic light-emitting diode (OLED) displays, small folding radius and low water vapor transmission rate(WVTR) are essential to ensure long lifetime of OLED device. For protect OLED from external moisture and oxygen, encapsulation is essential to OLED fabricating process. One of promising encapsulation methods is organic-inorganic multilayer thin film encapsulation(TFE) for flexible and stable OLED display. However, cracks can occur when cutting OLED panel with TFE. In this study, method for patterning inorganic layers has been investigated to fabricate sidewall patterned thin film encapsulation structure; Area selective atomic layer deposition, adhesion lithography. Although ASD process condition has been studied related to variables of ALD process; injection pressure of precursors, purge time, and chamber temperature, selectivity was not enough to apply to fabricate TFE. Al2O3 can be patterned through adhesion lithography which is one of patterning method using the difference in interfacial adhesion strength provided by depositing 1H,1H,2H,2H-Perfluorooctyltriethoxysilane (FOTS) SAM with shadow mask. Sidewall patterned TFE structure was fabricated with simple process comparing conventional photo lithography process by adhesion lithography for ALD deposited Al2O3 thin films and shadow mask for plasma polymer. 유기발광다이오드(OLED) 디스플레이의 장치의 수명을 연장하려면 접이 반경이 작고 수증기 투과율이 낮은 WVR(수증기 투과율)이 필수적이다. 따라서, 외부 수분 및 산소로부터 OLED를 보호하기 위해, 봉지 공정은 OLED 제조 공정에서 필수적이다. 유기-무기 다층 박막 봉지(TFE)공정은 유연하고 안정적인 OLED 디스 플레이를 위한 봉지 공정 중 하나이다. 단, TFE 방식으로 봉지한 OLED 패널을 절단하는 공정에서 균열이 발생할 수 있다. 이를 방지하기 위해 유기, 무기막이 모두 패턴된 TFE구조가 필요하다. 본 연구에서는 TFE구조 중, 무기층으로 사용되는 산화알루미늄(Al2O3) 박막을 패턴하는 방법을 연구하여 sidewall이 패터닝 된 TFE 구조를 제작하였다. 영역 선택적 원자층 증착법(ASD)과 접착력 리소그래피(adhesion lithography)를 통해 이를 구현하고자 하였다. TFE 제작에 사용된 유기막은 플라즈마 화학 기상 증착법(PECVD)을 통해 증착된 플라즈마 고분자 박막이며, 무기막은 원자층 증착법(ALD)을 통해 증착된 산화알루미늄(Al2O3) 박막이다. 영역 선택적 원자층 증착법 연구에서는 1H, 1H, 1H, 2H, 2HFerfluorooctylTriethyxilane(FOTS) 물질을 기상으로 자가조립막 처리하여 산화알루미늄의 원자층 증착을 방지하고자 했다. 공정 조건 중 전구체 주입 압력, 퍼지시간, 챔버 온도가 선택성에 미치는 영향을 연구하여 선택성을 개선하고자 했다. 이를 통해 35사이클까지 선택성을 유지하도록 개선되었으나, 이는 TFE를 위한 무기막 증착에 충분하지 않았다. 접착력 리소그래피 연구에서는 자가조립막 처리 유무에 따라 각 계면에서 접착력에 차이가 발생한다는 점을 이용해 산화알루미늄 박막을 패턴하고자 했다. 오인스-웬트법을 이용해 각 계면에서의 접착력을 이론적으로 계산하였다. FOTS 물질이 기상으로 자가조립막 된 표면에 원자층 증착법을 통해 전면으로 산화알루늄을 증착한 후 박리 공정을 통해 산화알루미늄 박막을 패터닝하였다. 접착력을 이용한 리소그래피 방식으로 원자층 증착법으로 증착된 산화알루미늄 박막을 패턴하고, 섀도우마스크를 이용한 플라즈마 화학 기상 증착법으로 플라즈마 고분자 박막을 패턴하여 TFE의 sidewall 구조를 구현하였다.

(The) effects of young children’s knowledge acquisition using tangible blocks

Cho, Jaewon Sungkyunkwan university 2016 국내석사

Korea and the major powers : an analysis of power structures in East Asia

Cho U.M.I. 1979 국내박사

Heterostuctured Surface Layers of Ni-based Cathode Materials for Li-ion Batteries

Yonghyun Cho Graduate School of UNIST 2013 국내박사

In an attempt to overcome the problems associated with LiNiO2, the solid solution series of lithium nickel-metal oxides, Li[Ni1-xMx]O2 (with M = Co, Mn, Al, Ti, Mg, etc.), have been investigated as favorable cathode materials for high-energy and high-power lithium-ion batteries. However, along with the improvement in the electrochemical properties in Ni-based cathode materials, the thermal stability has been a great concern, and thus violent reaction of the cathode with the electrolyte needs to be avoided. Firstly, we report a heterostructured Li[Ni0.54Co0.12Mn0.34]O2 cathode material which has a high energy and safety. The core part of the particle consist of LiNi0.54Co0.12Mn0.34]O2 with a layered phase (R3-m) and shell part with a thickness of < 0.5 μm consists of highly stable Li1+x[CoNixMn2-x]2O4 spinel phase (Fd-3m). The material demonstrates reversible capacity of 200 mAh g-1 and retains 95% capacity retention at the most severe test condition of 60 °C. In addition, amount of oxygen evolution from the lattice in the cathode with two heterostructures is reduced by 70%, compared to the reference sample. All these results suggest that the bulk Li[Ni0.54Co0.12Mn0.34]O2 consisting of two heterostructures satisfy the requirements for hybrid electric vehicles, power tools, and mobile electronics. Secondly, solid solution series of lithium nickel metal oxides, Li[Ni1-xMx]O2 (with M = Co, Mn, and Al) have been investigated intensively to enhance the inherent structural instability of LiNiO2. However, when a voltage range of Ni-based cathode materials was increased up to > 4.5 V, phase transitions occurring above 4.3 V resulted in accelerated formation of trigonal phase (P-3m1) and NiO phases, leading to and pulverization of the cathode during cycling at 60 °C. In an attempt to overcome these problems, LiNi0.62Co0.14Mn0.24O2 cathode material with pillar layers in which Ni2+ ions were resided in Li slabs near the surface having a thickness of ~10 nm was prepared using a PVPfunctionalized Mn precursor coating on Ni0.7Co0.15Mn0.15(OH)2. We confirmed the formation of a pillar layer via various analysis methods (XPS, HRTEM, and STEM). This material showed excellent structural stability due to a pillar layer, corresponding to 85 % capacity retention between 3.0 to 4.5 V at 60 °C after 100 cycles. In addition, amount of heat generation was decreased by 40 %, compared to LiNi0.70Co0.15Mn0.15O2.