list possible adverse effects in animals that might be seen from the proposed antigen or adjuvant injections and what measures will be taken should these adverse effects occur. (3) If feeder cells for supporting hybridoma colony growth will be collected from animals, describe the exact procedures that will be used to collect the feeder cells and the number of animals needed for this purpose. b. Consideration of Alternates to Animal Use for Collection of Antibody from Ascites. If animals will be used to expand hybridoma cell alternate research methods that can replace the use of animals must be considered. (1) Explain why cell culture systems for harvesting monoclonal antibodies are not in vitro adequate to meet the research objectives. (2) Provide the following information in tabular form: (a) Hybridoma cell line designation. (b) Number of animals used for ascites production. (c) Priming agent and volume. (d) Number and timing of priming injections. (e) Volume of injected hybridoma cells. (f) Number of abdominal taps before euthanasia. (3) Indicate what criteria will be used to determine if animals should be euthanatized prior to the last planned abdominal tap. c. Blood Collection as Part of Monoclonal Antibody Production. Indicate if survival blood collections will be obtained from animals following immunization or as a “pre-bleed” prior to immunization. (1) Provide the following table, including any prior to immunizations: “pre-bleeds” (a) Site of blood collection. (b) Amount of blood collected expressed as volume (ml) and percentage of body weight (assume 1 ml weighs 1 gram). (c) Number of blood collections. (d) Interval between collections. (2) Indicate if anesthetics, tranquilizers, or analgesics will be used prior to blood collection. If none will be used, justify the omission of these agents. Otherwise, provide the agent(s) including dose (mg per kilogram [kg]), volume (ml), route, and frequency and duration. d. Indicate if polyclonal antibodies will be produced as Polyclonal Antibody Production.
list possible adverse effects in animals that might be seen from the proposed antigen or adjuvant injections and what measures will be taken should these adverse effects occur. (3) If feeder cells for supporting hybridoma colony growth will be collected from animals, describe the exact procedures that will be used to collect the feeder cells and the number of animals needed for this purpose. b. Consideration of Alternates to Animal Use for Collection of Antibody from Ascites. If animals will be used to expand hybridoma cell alternate research methods that can replace the use of animals must be considered. (1) Explain why in vitro cell culture systems for harvesting monoclonal antibodies are not adequate to meet the research objectives. (2) Provide the following information in tabular form: (a) Hybridoma cell line designation. (b) Number of animals used for ascites production. (c) Priming agent and volume. (d) Number and timing of priming injections. (e) Volume of injected hybridoma cells. (f) Number of abdominal taps before euthanasia. (3) Indicate what criteria will be used to determine if animals should be euthanatized prior to the last planned abdominal tap. c. Blood Collection as Part of Monoclonal Antibody Production. Indicate if survival blood collections will be obtained from animals following immunization or as a “pre-bleed” prior to immunization. (1) Provide the following table, including any “pre-bleeds” prior to immunizations: (a) Site of blood collection. (b) Amount of blood collected expressed as volume (ml) and percentage of body weight (assume 1 ml weighs 1 gram). (c) Number of blood collections. (d) Interval between collections. (2) Indicate if anesthetics, tranquilizers, or analgesics will be used prior to blood collection. If none will be used, justify the omission of these agents. Otherwise, provide the agent(s) including dose (mg per kilogram [kg]), volume (ml), route, and frequency and duration. d. Polyclonal Antibody Production. Indicate if polyclonal antibodies will be produced as
SAS-SOP-2015-10-06.docx