The Application Protocol of CpG ODNs
-Edited by InvivoGen-
◎ Three Classes of CpG ODNs
CpG ODNs은 CpG motif에서 unmethylated CpG dinucleotide를 포함하는 짧은 single-stranded DNA 입니다. Toll-like receptor 9 (TLR9)에 의해 인식되며, 백신 보조제로 다양하게 연구되었습니다.
CpG ODNs은 구조 특성 및 활성에 기반하여 3가지로 분류됩니다: Class A (Type D), Class B (Type K), Class C
| Class | Structure | Applications |
| Class A | One PO central CpG-containing palindromic motif and one PS-modified 3’ poly-G string | - Stimulation of pDCs
- Induction of IFN-α production |
| Class B | A full PS backbone with one or more CpG dinucleotide | - Stimulation and proliferation of B cells
- NF-κB activation in TLR9-expressing recombinant cells
- Vaccine adjuvant |
| Class C | A complete PS backbone and a CpG-containing palindromic motif, combining the features of Class A and B CpG ODNs | - Combined applications of A-class and B-class CpG ODNs |
◎ Protocol in Mice Tumor Models
| Product | Mice Tumor Model | Protocol | PMID |
| ODN1668 | Colon Cancer | 20µg ODN1668 in 0.1ml PBS. Intratumor injection. | 29295954 |
| Application: TNFR2 신호전달 차단은 Colon Cancer mouse 모델에서 CpG ODN의 면역치료 효과 향상 |
ODN M362 | Hepatocellular Carcinoma | 5µg/mice of ODN M362. Intratumoral injection. | 25224571 |
| Application: TLR9 ligand CpGODN의 phosphorothioate 변형은 entry blockade에 의한 Hepatocellular의 poly(I:C)-induced apoptosis 억제 |
ODN2006 VacciGrade™, ODN2216,
ODN M362 | Breast Cancer | 100µg/mice MamA2.1 peptide and 100µg/mice ODNs. Intraperitoneal injection. | 31091800 |
| Application: ODN 2006 및 M362 oligodeoxynucleotide는 TLR9/-6 시너지 효과를 통해 강력한 보조제 효과를 발휘하여, Breast Cancer에 대한 Mammaglobin-A Peptide Specific Cytotoxic CD8+T Lymphocyte 반응 증가 |
VacciGrade™: High-quality pre-clinical grade
◎ Protocol in Animal Models of Infectious Diseases
| Product | Animal | Pathogen | Protocol | PMID |
ODN 1585,
ODN 1826,
ODN 2395,
ODN 2088 | Mice | HSV-1 | 50µg/mice of either ODNs. Intrenasal administration. | 23043942 |
| Application: Herpes simplex virus encephalitis mouse 모델에서 TLR9 반응 조절 |
| ODN2216 | Cotton rat | Measles Virus | 100µg/rat mixture of poly(I:C) and ODN 2216 (1:1). Subcutaneous injection. | 24262312 |
| Application: Neonatal cotton rat 샘플에서 TLR-3 및 TLR-9 agonist를 통한 interferon α의 유도는 Measles Virus에 대한 면역 반응을 자극 |
| ODN 2007 | Chicken | AIV H9N2 | 15µg inactivated H9N2 virus, 2-10µg ODN2007 and/or 0.4- 2µg Flagellin. Intramuscular injection. | 30222508 |
| Application: Chicken 샘플의 AIV H9N2 바이러스 백신의 보조제로서 TLR 5 및 TLR 21 비교 |
◎ Protocol in Animal Models of Other Diseases
| Product | Disease model | Protocol | PMID |
| ODN 1826 | Ischemic Brain Injury | 20 and 40μg/mice ODN1826. Intraperitoneal injection | 32510041 |
| Application: CpG-ODN1826을 사용하여 어린 수컷 mouse 샘플의 ischemic brain injury 감소: replication study |
ODN 2006,
ODN 1826 | Atherosclerosis | 2.5 - 120µg/mice ODN1826. Intravenous injection and subcutaneous injection. | 26751387 |
| Application: 고용량 CpG ODN 1826을 사용한 Toll-Like Receptor 9의 Proinflammatory Stimulation은 내피세포 재생을 저해하고 mouse 샘플의 Atherosclerosis을 촉진 |
| ODN 1668 | Heart Failure | 60µg/mice of ODN1668. Intraperitoneal injection. | 31312759 |
| TLR9 억제제 투여로 mouse 샘플의 Heart Failure 발병 및 진행 완화 |
▶ Related Products
더욱 다양한 CpG ODNs 제품 리스트 보기 (클릭)
◎ CpG ODN 1018 VacciGrade™
CpG 1018® 또는 1018 ISS라고도 알려진 CpG ODN 1018은 현재 FDA가 승인한 유일한 인간 백신용 CpG 기반 보조제입니다. ODN 1018은 대식세포 및 자연살해(NK) 세포로부터 IL-12, IL-18 및 IFN- γ의 생산, Th1 반응 촉진, 항체 생산 및 B 세포 증식에의 관여 등과 같은 면역학적 효과에 대해 광범위하게 테스트되었습니다.
이는 허가된 B형 간염 바이러스 백신 HEPLISAV-B®를 포함한 여러 승인된 백신의 구성 요소이기도 합니다.이는 HEPLISAV-B®를 포함한 여러 승인된 백신의 구성 요소이기도 합니다. 전염병(예: SARS CoV-2, 백일해) 또는 암에 대한 백신 전략에서 ODN 1018을 단독으로 또는 명반(alum)과 함께 사용하여 다양한 전임상 및 임상 시험이 진행 중입니다.
ODN은 amyloid pathology 제거 및 알츠하이머 치료제로서의 잠재력이 테스트되었습니다. 현재 CpG 1018®의 단독 투여가 초기 AD 환자에 미치는 영향을 조사하는 임상시험이 진행 중입니다. InvivoGen은 in vivo 사용에 적합한 고품질 pre-clinical grade의 ODN 1018 VacciGrade™*를 제공하며, 박테리아 오염 (lipoprotein 및 endotoxin) free가 확인되었습니다.
Influenza Virus Infection
Mice were primed and boosted intramuscularly (with a 3-week interval) with 3µg antigens and 3µg ODN 1018.
6 weeks after the prime, mice were intranasally challenged with 10x the 50% mouse lethal dose of the virus. |
Whooping Cough (Pertussis)
Mice were administered 50µl of vaccine intramuscularly, containing antigens (1/10-1/80 of human dose) and 10µg ODN 1018. Mice were boosted with the same vaccine formulations after 21d of the prime. |
Mesothelioma
Mice were peritumorally injected with 50µl endotoxin-free vaccine (50µg plasmid expressing CD40L, 25µg poly(I:C), and 25µg ODN 1018). The injections were repeated every other day for five doses. |
◎ Therapeutic Role of Free CpG ODNs
CpG ODNs는 선천성 (innate) 및 특이성 (specific) 면역의 강력한 활성화제로서, Th1 phenotype에 대한 면역반응을 유도합니다.Endosomal TLR9을 자극하여 Immune cascade를 유발함으로써, CpG ODNs는 암, 알레르기 및 감염성 질환 치료에 대한 효능을 조사했습니다.
주사 전 ODN을 mouse에 미리 처리하면 HSV-1 감염에 대한 생존율이 증가하며, 감염 후 ODN을 처리할 경우 염증 반응을 조절하는데 도움을 줍니다. CpG ODN 2006은 피부 T-세포 림프종 (Cutaneous T-cell lymphoma), 흑색종 (Melanoma) 및 신세포암종 (Renal cell carcinoma)을 포함한 다양한 종류의 암에서 항종양 효과가 광범위하게 연구되었습니다.
Herpes Simplex Virus Encephalitis(HSE)
The Pre-infection Studies: Mice were pre-treated with a single intranasal dose of 50 lg/mouse of either ligands (Poly(I:C), ODN 1585, ODN 1826, ODN 2395, ODN 2088) in a volume of 20µl of vehicle (0.9% saline). Then pretreated mice were infected intranasally one day later with 3800 PFUs of HSV-1 (herpes simplex virus type 1). Mice were sacrificed on day 3, 4, and 5 post-infection. The Post-infection Studies: Mice received the same treatment of ligands (Poly(I:C), ODN 1585, ODN 1826, ODN 2395, ODN 2088) 3 days following the infection with HSV-1 (4200 PFUs). Mice were sacrificed on day 3, 4, and 5 post-infection |
Colon Cancer
Mice were intraperitoneally injected with 100µg antibodies or mouse IgG1 in 0.2ml PBS at days 1, 5, and 9. At days 2 and 4, 20µg ODN1668 VacciGrade™* in 0.1ml PBS was intratumorally treated to the mice. One day after the last treatment, mice were sacrificed for harvesting tumor and lymphoid tissues. |
Alzheimer’s Disease (AD)
Aged monkeys were subcutaneously injected with 2mg/kg ODN 2006 (a month interval) for 24-month period. All injection were administered at 8-9 a.m. before the monkeys were fed. |
◎ Combination Therapy of CpG ODNs
보조제로서 항원과 CpG ODN의 조합은 별도 투여 시 보다 5배에서 500배까지 더 강한 therapeutic antigen-specific immune response를 유도할 수 있습니다. CpG ODN과 monoclonal 항체 치료 및 chemotherapy와 같은 다양한 치료 전략의 조합은 animal 모델에서 연구되었으며, 치료 효과가 입증되었습니다.
Lethal Influenza Pneumonia
Treatments were aerosolized to the animals for 30 min from a nebulizer driven by 10 L/min air supplemented. The following concentrations were used in the nebulizer reservoir: Pam2CSK4 10 µg/ml; poly(I∶C) 100 µg/ml; ODN2395 20 µg/ml. Ligand concentrations in the airway lining fluid are calculated to be Pam2CSK4 10ng/ml; poly(I∶C) 100ng/ml; ODN2395 20ng/ml. |
Avian Influenza Virus (AIV)
Chickens were vaccinated via intramuscular injection in the thigh muscle on days 7 and 21 post-hatch receiving 100 mL and 200 mL total volume, respectively. Chickens received 15 mg of formaldehyde inactivated H9N2 virus (around 960 HA units) per vaccination. Mixed with the inactivated H9N2 virus, chickens received either a high or low dose of FLA-ST Ultrapure* or CpG ODN 2007, a combination of both, or AddaVax. Serum and lachrymal secretions were collected from chickens weekly beginning 7 days post-primary vaccination (ppv) and ending 28 days ppv. |
Breast Cancer
Mice was injected with 100 µg/mice of endotoxin-free antigen into the peritoneal cavity with or without the ODN2006, ODN2216, ODN M362 (100 µg/mice for a given ODN type). |
◎ Novel Delivery System of CpG ODNs
안정성 (Stability), 약동학 (Pharmacokinetic) 및 생체분포 (Biodistribution)를 향상시키기 위해 nanoparticle, multi-component nanorods 및 liposome과 같은 새로운 전달 시스템이 개발되고 있습니다.
Delivered by PEGylated Nanoparticles
For intratumoral administration of immunostimulants, 0.2nmol of ODNs (ODN1585, ODN1826, ODN2395 and their controls) encapsulated within nanoparticles or unencapsulated were injected intratumorally at various time points. For intravenous therapeutic administration experiments, 1nmol of ODNs encapsulated within PEGylated nanoparticles or unencapsulated, in 150μL of PBS were injected into the lateral tail vein at various time points. |
Delivered by Alginate Coated Chitosan Nanoparticles
The vaccine contains suspension of alginate coated chitosan nanoparticles loaded with 10μg antigen and 10μg of the ODN1826 as adjuvant. 3 boosts within three weeks between each was administered in nasal mucosa. Four weeks after the final boost, mice were sacrificed. |
Mannose and CpG-ODN Coated Liposome Delivery
The liposomal vaccine was assembled by DC-targeting mannose and ODN 1826 on the surface of liposome loaded with anti-tumor agent. Mice were injected with 2μg assembled liposome in 100μl PBS on days 6, 9, 12, and 15 after the tumor implantation. |
Remakrs: 참조 자료의 모든 CpG-ODN 및 붉은색으로 표시된 제품은 InvivoGen에서 제공하고 있습니다.
*The marked products (ODN 1018 VacciGrade™, ODN1668 VacciGrade™, and FLA-ST Ultrapure) are exclusively provided by InvivoGen, as VacciGrade™ and Ultrapure grade.
▶ Related Products
*학술 문의 T. 031-728-3236, 3239 E. technical@dawinbio.com
*제품에 대한 보다 자세한 정보는 첨부파일을 통해 확인하실 수 있습니다.
#invivogen #다윈바이오 #ligand #agonist #TLR9 #CpG #ODN #vaccigrade
The Application Protocol of CpG ODNs
-Edited by InvivoGen-
◎ Three Classes of CpG ODNs
CpG ODNs은 CpG motif에서 unmethylated CpG dinucleotide를 포함하는 짧은 single-stranded DNA 입니다. Toll-like receptor 9 (TLR9)에 의해 인식되며, 백신 보조제로 다양하게 연구되었습니다.
CpG ODNs은 구조 특성 및 활성에 기반하여 3가지로 분류됩니다: Class A (Type D), Class B (Type K), Class C
- Induction of IFN-α production
- NF-κB activation in TLR9-expressing recombinant cells
- Vaccine adjuvant
◎ Protocol in Mice Tumor Models
ODN M362
ODN M362
VacciGrade™: High-quality pre-clinical grade
◎ Protocol in Animal Models of Infectious Diseases
ODN 1826,
ODN 2395,
ODN 2088
◎ Protocol in Animal Models of Other Diseases
ODN 1826
▶ Related Products
더욱 다양한 CpG ODNs 제품 리스트 보기 (클릭)
◎ CpG ODN 1018 VacciGrade™
CpG 1018® 또는 1018 ISS라고도 알려진 CpG ODN 1018은 현재 FDA가 승인한 유일한 인간 백신용 CpG 기반 보조제입니다. ODN 1018은 대식세포 및 자연살해(NK) 세포로부터 IL-12, IL-18 및 IFN- γ의 생산, Th1 반응 촉진, 항체 생산 및 B 세포 증식에의 관여 등과 같은 면역학적 효과에 대해 광범위하게 테스트되었습니다.
이는 허가된 B형 간염 바이러스 백신 HEPLISAV-B®를 포함한 여러 승인된 백신의 구성 요소이기도 합니다.이는 HEPLISAV-B®를 포함한 여러 승인된 백신의 구성 요소이기도 합니다. 전염병(예: SARS CoV-2, 백일해) 또는 암에 대한 백신 전략에서 ODN 1018을 단독으로 또는 명반(alum)과 함께 사용하여 다양한 전임상 및 임상 시험이 진행 중입니다.
ODN은 amyloid pathology 제거 및 알츠하이머 치료제로서의 잠재력이 테스트되었습니다. 현재 CpG 1018®의 단독 투여가 초기 AD 환자에 미치는 영향을 조사하는 임상시험이 진행 중입니다. InvivoGen은 in vivo 사용에 적합한 고품질 pre-clinical grade의 ODN 1018 VacciGrade™*를 제공하며, 박테리아 오염 (lipoprotein 및 endotoxin) free가 확인되었습니다.
Mice were primed and boosted intramuscularly (with a 3-week interval) with 3µg antigens and 3µg ODN 1018.
6 weeks after the prime, mice were intranasally challenged with 10x the 50% mouse lethal dose of the virus.
Mice were administered 50µl of vaccine intramuscularly, containing antigens (1/10-1/80 of human dose) and 10µg ODN 1018. Mice were boosted with the same vaccine formulations after 21d of the prime.
Mice were peritumorally injected with 50µl endotoxin-free vaccine (50µg plasmid expressing CD40L, 25µg poly(I:C), and 25µg ODN 1018). The injections were repeated every other day for five doses.
◎ Therapeutic Role of Free CpG ODNs
CpG ODNs는 선천성 (innate) 및 특이성 (specific) 면역의 강력한 활성화제로서, Th1 phenotype에 대한 면역반응을 유도합니다.Endosomal TLR9을 자극하여 Immune cascade를 유발함으로써, CpG ODNs는 암, 알레르기 및 감염성 질환 치료에 대한 효능을 조사했습니다.
주사 전 ODN을 mouse에 미리 처리하면 HSV-1 감염에 대한 생존율이 증가하며, 감염 후 ODN을 처리할 경우 염증 반응을 조절하는데 도움을 줍니다. CpG ODN 2006은 피부 T-세포 림프종 (Cutaneous T-cell lymphoma), 흑색종 (Melanoma) 및 신세포암종 (Renal cell carcinoma)을 포함한 다양한 종류의 암에서 항종양 효과가 광범위하게 연구되었습니다.
The Pre-infection Studies: Mice were pre-treated with a single intranasal dose of 50 lg/mouse of either ligands (Poly(I:C), ODN 1585, ODN 1826, ODN 2395, ODN 2088) in a volume of 20µl of vehicle (0.9% saline). Then pretreated mice were infected intranasally one day later with 3800 PFUs of HSV-1 (herpes simplex virus type 1). Mice were sacrificed on day 3, 4, and 5 post-infection. The Post-infection Studies: Mice received the same treatment of ligands (Poly(I:C), ODN 1585, ODN 1826, ODN 2395, ODN 2088) 3 days following the infection with HSV-1 (4200 PFUs). Mice were sacrificed on day 3, 4, and 5 post-infection
Mice were intraperitoneally injected with 100µg antibodies or mouse IgG1 in 0.2ml PBS at days 1, 5, and 9. At days 2 and 4, 20µg ODN1668 VacciGrade™* in 0.1ml PBS was intratumorally treated to the mice. One day after the last treatment, mice were sacrificed for harvesting tumor and lymphoid tissues.
Aged monkeys were subcutaneously injected with 2mg/kg ODN 2006 (a month interval) for 24-month period. All injection were administered at 8-9 a.m. before the monkeys were fed.
◎ Combination Therapy of CpG ODNs
보조제로서 항원과 CpG ODN의 조합은 별도 투여 시 보다 5배에서 500배까지 더 강한 therapeutic antigen-specific immune response를 유도할 수 있습니다. CpG ODN과 monoclonal 항체 치료 및 chemotherapy와 같은 다양한 치료 전략의 조합은 animal 모델에서 연구되었으며, 치료 효과가 입증되었습니다.
Treatments were aerosolized to the animals for 30 min from a nebulizer driven by 10 L/min air supplemented. The following concentrations were used in the nebulizer reservoir: Pam2CSK4 10 µg/ml; poly(I∶C) 100 µg/ml; ODN2395 20 µg/ml. Ligand concentrations in the airway lining fluid are calculated to be Pam2CSK4 10ng/ml; poly(I∶C) 100ng/ml; ODN2395 20ng/ml.
Chickens were vaccinated via intramuscular injection in the thigh muscle on days 7 and 21 post-hatch receiving 100 mL and 200 mL total volume, respectively. Chickens received 15 mg of formaldehyde inactivated H9N2 virus (around 960 HA units) per vaccination. Mixed with the inactivated H9N2 virus, chickens received either a high or low dose of FLA-ST Ultrapure* or CpG ODN 2007, a combination of both, or AddaVax. Serum and lachrymal secretions were collected from chickens weekly beginning 7 days post-primary vaccination (ppv) and ending 28 days ppv.
Mice was injected with 100 µg/mice of endotoxin-free antigen into the peritoneal cavity with or without the ODN2006, ODN2216, ODN M362 (100 µg/mice for a given ODN type).
◎ Novel Delivery System of CpG ODNs
안정성 (Stability), 약동학 (Pharmacokinetic) 및 생체분포 (Biodistribution)를 향상시키기 위해 nanoparticle, multi-component nanorods 및 liposome과 같은 새로운 전달 시스템이 개발되고 있습니다.
For intratumoral administration of immunostimulants, 0.2nmol of ODNs (ODN1585, ODN1826, ODN2395 and their controls) encapsulated within nanoparticles or unencapsulated were injected intratumorally at various time points. For intravenous therapeutic administration experiments, 1nmol of ODNs encapsulated within PEGylated nanoparticles or unencapsulated, in 150μL of PBS were injected into the lateral tail vein at various time points.
The vaccine contains suspension of alginate coated chitosan nanoparticles loaded with 10μg antigen and 10μg of the ODN1826 as adjuvant. 3 boosts within three weeks between each was administered in nasal mucosa. Four weeks after the final boost, mice were sacrificed.
The liposomal vaccine was assembled by DC-targeting mannose and ODN 1826 on the surface of liposome loaded with anti-tumor agent. Mice were injected with 2μg assembled liposome in 100μl PBS on days 6, 9, 12, and 15 after the tumor implantation.
Remakrs: 참조 자료의 모든 CpG-ODN 및 붉은색으로 표시된 제품은 InvivoGen에서 제공하고 있습니다.
*The marked products (ODN 1018 VacciGrade™, ODN1668 VacciGrade™, and FLA-ST Ultrapure) are exclusively provided by InvivoGen, as VacciGrade™ and Ultrapure grade.
▶ Related Products
*학술 문의 T. 031-728-3236, 3239 E. technical@dawinbio.com
*제품에 대한 보다 자세한 정보는 첨부파일을 통해 확인하실 수 있습니다.
#invivogen #다윈바이오 #ligand #agonist #TLR9 #CpG #ODN #vaccigrade