Monoclonal Antibodies

Presentation on theme: "Monoclonal Antibodies"— Presentation transcript:

1 Monoclonal Antibodies
For Cancer Therapeutics LG생명과학연구소 박영우

2

3 C C2 C3 C1 C C2 C3 C1 C1 C 항체 CDR Variable (V) scFv Constant region
Fab 생물체는 외부로부터 침입해 오는 병원균을 특이적으로 인식하여 이에 대응하는 체계로 면역계를 발달시켜왔음 이처럼 외부로부터 병원체 등 이물질이 들어오면 이것에 대항하는 물질이 만들어지는데 이를 항체라 한다. 항체는 4개의 폴리펩타이드로 되어 있는데 2개의 길다란 H사슬과 2개의 짧은 L 사슬이 disulfide bond로 연결되어 Y 모양을 하고 있음. 각 사슬의 말단에 있는 변이부는 항원을 찾아 결합하는 부위로 항체마다 다름. 변이부 아래에는 같은 부류에 속하는 항체끼리는 똑같은 불변부가 있음 항체의 생성 과정:항원이 몸에 들어오면 B세포는 항원을 인식하고 항원과 결합하고 이어서 활발한 세포분열을 일으켜 그 수를 늘린다. 그 뒤 늘어난 림프구는 그 항원에 대한 항체를 생성할 수 있는 형질세포로 분화되어 다량의 항체를 생성함 항체는 특정한 항원하고만 결합하는 특이성이 있는데 이를 항원-항체 반응의 특이성이라 함.

4 Specific Targeting Macrophage Attack (ADCC) Complement Attack (CDC) Stability IgG1: 23 days New Function

5 Therapeutic Effect of Monoclonal Antibody
Blocking cell activity and immune functions Activating cell activity and immune functions Delivering therapeutic agents

6 A B C D ANTIBODY의 작용 기작 Neutralizing antibody
타겥 분자를 inactivation 시킴 (Antagonistic Ab) 타겥 분자를 activation 시킴 (Agonistic Ab) A host immune system Killing effect (Herceptin, Retuxan) B Delivery Isotope Toxin Chemicals Cytokine (ex. IL2 등) C Cytokine 의 활성억제 및 제거 (Remicade, Xolair, Anti-VEGF) D

7 Technology Improvement
From Mouse Human Ab From Human Chimeric Ab Humanized Ab Mouse Ab Mouse Ab Human Ab

8 항체 제조 핵심기술 보유회사 HYBRIDOMA (Standard) 면역 (쥐) B CELL mAb (mouse) HYBRID Mouse mAb Humanize 면역 (쥐) B CELL mAb (mouse) CDR graft (Modeling) HYBRID Transgenic Mouse 면역 (쥐) B CELL mAb (human) HYBRID Human Naive LIBRARY mAb (human)

9 시장 및 개발현황 Anti-EGFr Anti-VEGF Anti-cancer mab HUMIRA Affinity increase
2001 2000 1998 1997 1995 1994 1986 1985 1975 2002 2003 HUMIRA Affinity increase Yeast Display Fully Human Phage Display RA Hybridoma Panorex OKT3 Zevalin Zenapax Herceptin Synagis Mylotarg Campath Humanization Phage Display Transgenic Mice Yeast Display Rituxan Remicade Simulect Chimerization ReoPro

10 FUTURE TREND FULLY HUMAN ANTIBODY

11 HIGH QUALITY, ULTRADIVERSITY GENE POOL
LG ANTIBODY LIBRARY HIGH QUALITY, ULTRADIVERSITY GENE POOL 100,000,000,000 TRANSFORMANTS -ⅱ-

12 HIGH QUALITY 1011 different phage pool
scFv HIGH QUALITY 1011 different phage pool

13 PANNING OPGL

14 BINDING OPGL

15 WASHING OPGL

16 ELUTION OPGL

17 AMPLIFICATION

18 OPGL MYC control ELISA signal Panning OPGL PANNING RESULT 1st 2nd 3rd
3.50 3.00 2.50 ELISA signal 2.00 1.50 1.00 0.50 0.00 1st 2nd 3rd 4th Panning

19 INDIVIDUAL CLONE ANALYSIS (4TH PANNING)
Fully Human Anti-OPGL scFv fragment 6.000 5.000 4.000 Hybridoma clones 3.000 2.000 ~109 nM 1.000 0.000 #1 #3 #8 #9 #12 #21 #22 #23 2E2 7A2

20 Anti Cancer Targets

21 Targets for Monoclonal Antibodies
Receptor Ligand Secreted Regulatory Proteins 인체는 수많은 세포로 이루어져있으며, 이들은 세포와 또 다른 세포사이의 조화로운 networking에 의해서 유지된다. 이러한 networking은 세포표면의 각종수용체 (receptor)와 리간드 (ligand) 및 분비 단백질 (예 홀몬, 싸이토카인 등)에 의해 이루어지고있으며, 이러한 조화가 깨질 경우 우리는 각종질환에 걸리게 되는 것이다. 따라서 수용체, 리간드 및 분비되는 단백질들은 의약품개발에 좋은 타겥이 된다.

22 Genome-Wide Expression Profiling
Target Discovery by Genome-Wide Expression Profiling

23 Target Discovery by Expression Profiling
Tissue Samples Expression Database Data Analysis Gene List Target Prioritization Expression Validation Target Validation

24 Affymetrix GeneChip Technology
Human Genome U133                                                                               Probe Set PM MM Probe pair Cell : perfectly complementary to a target sequence : identical to PM except for a single base mismatch in its center * The difference in hybridization signals between PM and MM serve as indicators of specific target abundance.

25 LG GI Cancer Database Stomach Liver Pancreas
 LGLS Proprietary Gastrointestinal Tract Cancer Genome DB (106 tissue samples). Clinical Samples GeneChip Analysis LG GI Cancer Database Stomach Liver Pancreas Normal Hyperplastic Polyp Epithelial Dysplasia Early Gastric Cancer Advanced Gastric Cancer Normal Chronic Hepatitis Liver Cirrhosis Hepatocellular Carcinoma IH Cholangiocarcinoma GB Cancer Normal Chronic Pancreatitis Adenocarcinoma IPMT Solid Papillary Neoplasm

26 PCA of Liver Samples Normal Hepatocellular Carcinoma Chronic Hepatitis
Liver Cirrhosis

27 PCA of Stomach Samples Normal Adenocarcinoma Chronic inflammation
Epithelial dysplasia Signet ring cell carcinoma Hyperplastic polyp

28 Novel Genes Associated with Cancers
LBFL109 : GTPase activating protein LBFL110 : Receptor LBFL123 : Transporter LBFL131 : Membrane protein LBFL164 : Kinase LBFL167 : Intracellular signaling molecule LBFL169 : Transporter LBFL301 : Secreted protein LBFL302 : Transcription factor LBFL303 : Transporter LBFL304 : Protease ? LBFL305 : Kinase LBFL306 : ANK repeat LBFL307 : Membrane protein LBFL308 : Transferase LBFL309 : Membrane protein LBFL310 : Membrane protein LBFL312 : Similar to rat embigin (proliferation / differentiation) LBFL313 : Secreted protein LBFL314 : Calpain homolog (migration / proliferation) LBFL315 : Isomerase

29  Human Cancer Genome DB
 Processed DB of Gene Logic’s Oncology Datasuite (1,094 tissue samples). Breast Cervix Colon Duodenum Endometrium Esophagus Kidney Liver Lung Lymph Node Myometrium Ovary Pancreas Prostate Rectum Skin Stomach Oncology Datasuite*  GeneChip Analysis Normal vs Cancer Data Mining Cancer Related Genes

30 Tissue samples used in Gene Logic’s Oncology Datasuite

31 Immunotoxin approach to anti-IL2RA in HD
IL2R (LYMPHNODE) Immunotoxin approach to anti-IL2RA in HD IL2RG BREAST DUOD KIDNEY LYMPH PANC CERVIX COLON ENDOM ESOPH LIVER LUNG MYOM OVARY PROST RECTUM SKIN STOMACH IL2RA IL2RB

32 KIDNEY Clear Cell & Renal Cell Carcinoma specific
CCA OC RCC N TNFAIPx Clear Cell Carcinoma Renal Cell Carcinoma BREAST DUOD KIDNEY LYMPH PANC CERVIX COLON ENDOM ESOPH LIVER LUNG MYOM OVARY PROST RECTUM SKIN STOMACH GPRxx

33 KIDNEY Oncocytoma specific
CCA OC RCC N gene 1 BREAST DUOD KIDNEY LYMPH PANC CERVIX COLON ENDOM ESOPH LIVER LUNG MYOM OVARY PROST RECTUM SKIN STOMACH gene 2 gene 3

34 CXCR family BREAST DUOD KIDNEY LYMPHNODE PANCREASE IDL N ID IL CCA OC
RCA N HD N ML N AC N AC BREAST CERVIX DUOD ESOPH KIDNEY LUNG LYMPH OVARY PANC RECTUM SKIN STOMACH COLON ENDOM LIVER MYOM PROST

35 GENE1 (BREAST)

36 Related Platform Technology
LG ANTIBODY LIBRARY & Related Platform Technology LG GI Cancer Database Gene Logic’s Oncology Datasuite Bioinformatics Target Selection & Validation Collaboration 서울대 암연구소 유전체 사업단 Therapeutic and diagnostic Antibody