Specific Toxicity Studies

Local Irritation Studies Local irritation studies are conducted in order to determine the irritancy of pharmaceuticals, medical devices, chemical compounds, and agrochemicals to skin and muscle tissue, blood vessels, and mucosal tissue. From approval of the protocol until submission of the draft report takes 2 to 2.5 months. If pathological examinations are included, the period is 3.5 to 4.0 months. Primary skin irritation Intracutaneous irritation Primary eye irritation Blood vessel irritation Intramuscular/subcutaneous local irritation Cumulative skin irritation Phototoxicity (in vivo, in vitro) Cumulative eye irritation (including contact lens application studies) Both primary and cumulative irritation studies can encompass nasal, vaginal, rectal, and oral mucosa irritancy.
Antigenicity/Sensitization Studies Antigenicity/sensitization studies are conducted to investigate the inducibility of immediate and delayed-type allergies after exposure to pharmaceuticals, medical devices, or agrochemicals. ( ): Time from approval of the protocol until submisssion of the draft report. Skin sensitization studies Maximization method (3 months) Adjuvant and patch method (3 months) Buehler method (2.5 months) Skin photosensitization studies Adjuvant and strip method (2.5 months) Antigenicity studies Active systemic anaphylactic response (3 months) EPassive cutaneous anaphylactic response (3 months) Cutaneous response (3 months) Antibody titer in serum by ELISA (1.5 months, 3.5 months including preparation of serum)
Ocular Toxicity Studies SNBL has extensive experience in a wide range of ocular toxicity studies using rabbits, non-human primates, and dogs. These studies require approximately two months, but the duration may vary depending on the protocol.	Photo of fluorescein-stained fundus of cynomolgus monkey
Macroscopic examination by Draize method Pupillary response to several drugs Pupillary reflex examination [after-dark adaptation, under scattered light, and light irradiation] Examination of anterior portion of the eye with a slit lamp (corneal damage examination using fluorescein) Fluorescein ocular fundus examination Intra-ocular pressure examination Refractometer examination Examinations of optic media and ocular fundus using binocular indirect ophthalmoscope, slit lamp, and fundus camera Electroretinogram examination (a- and b-wave, oscillatory potential, reaction in cone and rod cells, and flicker response)
Biological Safety Studies for Medical Devices SNBL offers the following studies in compliance with GLP: Cytotoxicity (colony forming inhibition) Sensitivity Genotoxicity Irritation General toxicity Pyrogenicity Blood compatibility (hemolytic toxicity study) Implantation studies in beagles and rabbits (bone/muscle) Extraction prerequisites can be investigated in non-GLP studies.	Bone implantation, decalcification, paraffin sectioning, H.E. Staining

Mutagenicity SNBL has conducted more than 1,000 medical device, foodstuff, and OECD-guideline compliant studies. Bacterial reverse mutation test (reporting time: 1.5 months) Chromosomal aberration test using cultured mammalian cells or human lymphocytes (2 months) Micronucleus test using bone marrow or peripheral blood (2 months) Mouse lymphoma assay (2.5 months) Comet assay (2 months)	Bone implantation, electropolish, SEM photograph

Immunotoxicity Studies Typical immunotoxicity tests and parameters Hematology: Total and absolute leukocyte counts (standard parameter) Blood chemistry: Globulin concentration (including γ-globulin), A/G ratio, and non-human primate IgG, IgM, and IgA concentrations Gross pathology: Lymphatic organs and tissue Organ weights: Thymus and spleen (lymph node) Histopathology: Thymus, spleen, lymph nodes, bone marrow, Payer's patches, BALT, NALT, and CALT Immunophenotyping is also offered. Immunotoxicity studies T-cell dependent antibody response (TDAR) Induced by Keyhole limpet hemocyanin, a T-cell dependent antigen, in cynomolgus monkeys and rats Antibody titer measurement by ELISA Immunophenotyping Peripheral blood: T-cell (helper, killer), B cell, NK cell Spleen, thymus, and bone marrow	Comet Assay
<Standard parameters> Cynomolgus monkeys: CD3⁺CD45⁺, CD3⁺CD4⁺CD45⁺, CD3⁺CD8⁺CD45⁺, CD3^-CD20⁺CD45⁺, CD3^-CD16⁺CD45⁺ Rats: CD3⁺, CD3⁺CD4⁺CD8^-, CD3⁺CD4^-CD8⁺, CD3^-CD45RA⁺, CD3^-NKR-P1A⁺ Mice: CD3e⁺, CD45R/B220⁺, CD3e⁺CD4⁺CD8a^-, CD3e⁺CD4^-CD8a⁺, CD49b/Pan-NK cells⁺CD3e^- Measured by FACS Calibur (BD Bioscience) four color system Prior validation is required depending on the combination of parameters. Other parameters may also be possible after validation Immunohistochemical examination (thymus, spleen, lymph nodes, etc) <Measurable parameters> Cynomolgus monkey: CD3, CD4, CD8, CD13, CD16, CD20, CD68, CD83 Rat: CD3, CD8, PECAM-1(CD31), CD45RA, Pan-B, ED-1, Ki-M2R NK activity examination (cynomolgus monkey) NK activity examination is measured by Flow cytometry targeting K562 cells. Cell-mediated immunity examination Cynomolgus monkeys are sensitized with tetanus toxoid, and challenged intradermally to cause a delayed type hypersensitivity (DTH) reaction a few weeks later. Immunosuppressive efficacy of test article can be evaluated. Other examinations Blood cytokine measurement (monkey: IL-2, IL-4, IL-5, IL-6, IFN-γ, TNF), peripheral blood cell blastogenesis, etc.

Antibody Production and Measurement Antibody production triggered by biodrugs (including human antibodies) Polyclonal antibody production in non-human primates, rabbits, rats, and mice triggered by drugs, medical products, chemical substances, or agrochemicals Antibody purification and labeling Antibody purification with Protein A and Protein G from serum, ascites, and culture supernatant Antibody labeling: Antibody and enzyme (horseradish peroxidase), FITC, and biotin Idiotype human antibody purification Development and validation of measurement methods GLP-compliant methods of measurement of biotechnology-based drug concentrations have been developed and validated. Others Purification of proteins and hormones, measurement of enzyme activity, cytotoxicity studies, drug-efficacy screening studies using cells, flow cytometry, and other in vitro studies. Biacore T100 is now in use