NIH Grant Application, Vertebrate Animals Section

GenScript is dedicated to ensuring the welfare of animals that make scientific discovery possible. With an Office of Laboratory Animal Welfare (OLAW) Assurance, Institutional Animal Care and Use Committee (IACUC), and AAALAC accreditation, GenScript has established and will maintain a program for activities involving animals in accordance with the "Guide for the Care and Use of Laboratory Animals", PHS Policy for all animals, and the regulations of local government.

• Chinese government animal use license #'s: SYXK(SU)2012‐0053 and SYXK(SU)2012‐0054)
• GenScript's OLAW Assurance #: A5892-01

When filling out grant applications for the national institute of health (NIH), studies which involve the use of vertebrate animals are required to provide specifications for the following requests. The answers provided pertain to our general custom antibody generation services and may be further customized to fit the customers needs where appropriate.

Answers to NIH Requests

• Descriptions of Procedures

  Please refer to yo ur specific protocol provided for exact numbers. Typical specifications for animals used in antibody generation are listed in the table below:

  
  

  Service	Age or Weight	Species and Strain	Sex
  Antibody Production	7 weeks	BALB/c mouse	Female
  	180-200 g	Wistar rat	Female
  	1.8-2.5 kg	New Zealand Rabbit	Male

  Antibody development involves the use of animals as hosts to generate an immune response against the antigen target. Host animals are pre-bled via the orbital sinus (rodents) or the marginal ear vein (rabbits) in order to acquire pre-immune serum to be 50-100 μl/mouse or rat and 0.5 ml/rabbit. Typical immunization involves subcutaneous injection with an emulsion of incomplete Freund's adjuvant (IFA) and target protein or peptide. Response to immunization is measured via orbital sinus (rodents) or marginal ear vein (rabbits) bleed samples. Subsequent exposures to antigen via subcutaneous injection are repeated every other week until sufficient test titers are reached. Serum (polyclonal antibody) or lymphocyte (monoclonal antibody) harvest is performed via terminal procedure including deep anesthesia and immediate euthanasia. Serum is collected through cardiac puncture, and lymphocytes are collected from spleen or local lymph nodes at this time.

  Generation of antibodies requires a mammalian immune system in order to generate a full repertoire of antibodies. Mice, rats, and rabbits are well studied, accepted, and proven hosts for generating useful antibodies for both research and therapeutic or industrial applications. Use of multiple animals is necessitated in order to generate required volumes of material produced (polyclonal serum) and/or provide sufficient statistical power to elicit an appropriate immune response (monoclonal or polyclonal).

• Justifications

  Generation of antibodies requires a mammalian immune system in order to generate a full repertoire of antibodies. Mice and rabbits are well studied, accepted, and proven hosts for generating useful antibodies for both research and therapeutic or industrial applications. Use of multiple animals is necessitated in order to generate required volumes of material produced (polyclonal serum) and/or provide sufficient statistical power to elicit an appropriate immune response (monoclonal or polyclonal).

• Minimization of Pain and Distress

  Whenever possible, we strive to harmonize scientific requirements with animal well‐being issues and, when not possible, ensure that the animal research team has a clear understanding of the issues that warrant an exception. We ensure that the animals are monitored closely for unexpected outcomes not delineated in the approved animal study protocol and report any such outcomes to the veterinarian and IACUC. Animals are observed for obvious distress or discomfort, rapid or labored breathing, pallor, hunched posture, inactivity, dehydration, weight loss, roughened hair coat and ambulation difficulty.

  More subtle behavioral indices are needed to provide an indication whether the animals are experiencing mild to moderate pain or distress. Such subtle methods might include observing an animal under red light. Rodents are generally more active under red light, and declines in activity will be easier to identify than when the animals are relatively quiescent under bright light.

  Circumstances when animals may experience discomfort, distress, pain or injury

  Routine procedures such as injections and blood sampling from veins that produce only mild, transient pain, or discomfort. Another example is an observational study of animal behavior.

  Veterinarians review each research protocol prior to IACUC review and are available for consultation during the protocol submission process and an "as needed basis". Veterinarians advise animal users on drug selection and proper usage. Non-pharmacologic means is not used to moderate pain. IACUC approved anesthesia agents is sodium pentobarbital.

  Induction Agent	Species	Dose Rang(mg/kg)	Route of Administration
  Sodium Pentobarbital	Mice	30-90	IP
  	Rats	30-60	IP
  	Rabbits	40	IP

  IACUC approved analgesia agents list

  Agents	Species	Dose Range (mg/kg)	Route of Administration
  Acetominophen (Tylenol)	Rat	50	SC, IP
  	Rat	100	PO
  	Rodent	110-305	PO, IP
  Aspirin	Rat	100	PO
  	Rodent	20	SC
  	Rodent	100-120	IP
  Carprofen	Rat	5-15	SC
  Meloxicam	Mouse	1-2	PO, SC
  	Rat	1-2	PO, SC
  	Rabbit	0.3	PO, SC
  Flunixin	Mouse	1-2.5	SC
  	Rat	1-2.5	SC
  	Rabbit	1	SC

• Veterinary Care

  Provisions for special care or housing
  Room temperature and humidity are observed and recorded daily on a high/low thermometer and hygrometer located in each room. Observations are recorded on individual room maintenance sheets by the husbandry staff. Current room ventilation data is provided in Appendix (HVAC data summary). In case of temperature and humidity fluctuation beyond the range set in the SOP, the husbandry staff will investigate it and take immediate proper action. According the Guide, temperature set points in rodent facility is 22±2 ℃, humidity set points is 50±20%. Temperature set points in rabbit and chicken facility is 20±2 ℃, humidity set points is 50±20%.

  Plans for post-surgical care, if applicable
  The post‐operative care will be provided by the veterinary staff of the animal facility, where the animal will be housed. This includes monitoring, record keeping, and treatments. Treatment will be provided according to post‐operative orders agreed to by the Principal Investigator.

• Euthanasia, humane experimental endpoints, if relevant

  An animal in chronic pain or distress that cannot be relieved by analgesics, will be euthanized unless prior scientific justification for these humane endpoints are approved by the IACUC. Rodents are primarily euthanatized by CO₂ gas or cervical dislocation. Rabbits are euthanatized using pentobarbital‐based products (Sodium pentobarbital). An American Veterinary Medical Association (AVMA) approved method of euthanasia will be used. Any exception to the AVMA euthanasia recommendations must be described in the IACUC protocol and specifically approved by the IACUC.

Antibody Generation Related Services

Looking for a cost effective way to spend your grant? GenScript's antibody services provide the best value, guarantees, and turn around time to expedite your research. Just a few of our antibody generation services include:

• PolyExpress™ Antibody Service: our polyclonal antibody service can deliver you 3 antibodies in just 45 days starting from only your sequence.
• MonoExpress™ Antibody Service: high quality, purified monoclonal antibodies guaranteed to work in your application.
• Semi-custom Monoclonal Antibody Production Services: you can go from antigen sequence to antibody production with as little as 3 easy decisions.

Our customer service representatives are available 24 hours a day, Monday through Friday, to assist you.