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1. Monoclonal Antibody Production: Will monoclonal antibodies be produced in animals or harvested from hybridoma cell lines as part of this project?
FORMCHECKBOX No. Proceed to item 3.
FORMCHECKBOX Yes. Complete item 1.a.
a. Is antibody harvest limited to existing hybridoma cell lines with no further immunizations or lymphocyte fusions planned?
FORMCHECKBOX Yes. Proceed to item 2 below.
FORMCHECKBOX No. Fill out items 1.b and 1.c below; then proceed to item 2.
b. Complete the following table regarding the immunization protocol for the animals prior to lymphocyte harvest for hybridoma creation. For each antigen for which multiple immunization days will be used, use a separate row in the table for each immunization day.
TABLE 1 MONOCLONAL ANTIBODYInjection day (e.g. day 0, 7, 30, etc.)AntigenAdjuvantAmount antigenTotal injection volume Number of divided injections Injection route and anatomical location
(e.g., SC, dorsum) F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m g F O R M T E X T m L F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m g F O R M T E X T m L F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m g F O R M T E X T m L F O R M T E X T F O R M T E X T
c . I f f e e d e r c e l l s f o r s u p p o r t i n g h y b r i d o m a c o l o n y g r o w t h w i l l b e c o l l e c t e d f r o m a n i m a l s , d e s c r i b e t h e e x a c t p r o c e d u r e s t h a t w i l l b e u s e d t o c o l l e c t t h e f e e d e r c e l l s a n d t h e n u m b e r o f a n i m a l s n e e d e d f o r t h i s p u r p o s e .
F O R M T E X T
2 . A l t e r n a t i v e s : C o n s i d e r a l t e r n a t e r e s e a r c h m e t h o d s t h a t c a n r e p l a c e t h e u s e o f a n i mals. Will any animals be used to expand hybridoma cell lines so that antibody can be harvested from ascites fluid?
FORMCHECKBOX No. Proceed to item 2.d.
FORMCHECKBOX Yes. Complete items 2.a-2.c below; then proceed to item 2.d.
a. Explain why in vitro cell culture systems for harvesting monoclonal antibodies are not adequate to meet the research objectives.
FORMTEXT
b . C o m p l e t e t h e f o l l o w i n g t a b l e .
T A B L E 2 H Y B R I D O M A C E L L L I N E S H y b r i d o m a c e l l l i n e d e s i g n a t i o n N u m b e r o f a n i m a l s u s e d f o r a s c i t e s p r o d u c t i o n P r i m i n g a g e n t a n d v o l u m e ( m L ) N u m b e r a n d t i m i n g o f p r i m i n g i n j e c t i o n s V o l u m e o f i n j e c t e d h y b r i d o m a c e l l s ( m L ) N u m b e r o f a b d o m i n a l t a p s b e f o r e e u t h a n a s i a F O R M T E X T F O R M T E X T F O R M T E X T m L F O R M T E X T F O R M T E X T m L F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m L F O R M T E X T F O R M T E X T m L F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m L F O R M T E X T F O R M T E X T m L F O R M T E X T
c . E u t h a n a s i a : W h a t c r i t e r i a w i l l b e u s e d t o d e t e r m i n e t h e t e r m i n a l a b d o m i n a l t a p ?
F O R M T E X T
d . B l o o d c o l l e c t i o n : W i l l s u r v i v a l b l o o d c o l l e c t i o n s b e o b t a i n e d f r o m a n i m a l s f o l l o w i n g i m m u n i z a t i o n o r a s a p r e - b l e e d p r i o r t o i m m u n i z a t i o n ?
F O R M C H E C K B O X N o . P r o c e e d t o i t e m 3 .
F O R M C H E C K B O X Y e s . Complete items i) and ii) below; then proceed to item 3.
i) Complete the following table; include any pre-bleeds prior to immunizations.
TABLE 3 BLOOD COLLECTIONSite of blood collection*Amount of blood collected expressed as volume (mL)/ and % of body weight (assume 1 ml = 1 gram)Number of blood collectionsInterval between collections FORMTEXT F O R M T E X T m L / F O R M T E X T % F O R M T E X T F O R M T E X T w e e k s F O R M T E X T F O R M T E X T m L / F O R M T E X T % F O R M T E X T F O R M T E X T w e e k s F O R M T E X T F O R M T E X T m L / F O R M T E X T % F O R M T E X T F O R M T E X T w e e k s
* T o a p p r o x i m a t e a c c e p t a b l e b l o o d c o l l e c t i o n v o l u m e s , a s s u m e t h e t o t a l b l o o d v o l u m e o f a h e a l t h y a n i m a l a s 7 % o f i t s b o d y w e i g h t , a n d t h a t 1 0 % o f t h e t o t a l b l o o d v o l u m e c a n b e c o l l e c t ed every 2 weeks without causing significant physiological change. (Example: acceptable blood volume for a 30 g mouse ( 0.07 X 30 g X 0.10 = .21 mL.) Blood volumes in excess of 10% will be considered with scientific justification. Typically the maximum blood available for a terminal procedure is 50% of blood volume MUST be taken with the animal under anesthesia.
ii) Will anesthetics, tranquilizers, or analgesics be used prior to blood collection?
FORMCHECKBOX No. Justify the omission of pain-r e l i e v i n g a g e n t s b e l o w ; t h e n p r o c e e d t o i t e m 3 .
F O R M C H E C K B O X Y e s . D e s c r i b e t h e a d m i n i s t r a t i o n o f p a i n - r e l i e v i n g a g e n t s i n c l u d i n g d o s e ( m g / k g ) , v o l u m e ( m L ) , r o u t e , a n d f r e q u e n c y / d u r a t i o n ; t h e n p r o c e e d t o i t e m 3 .
F O R M T E X T
3 . P o l y c l o n a l Antibody Production: Will polyclonal antibodies be produced in this species of animal as a part of this project?
FORMCHECKBOX No. Do not complete items 3.a.-3.c. Go to item 4 below.
FORMCHECKBOX Yes. Complete items 3.a.-3.c., then go to item 4.
Complete the following table. For each antigen for which multiple immunization days will be used. Use a separate row in the table for each day.
TABLE 4 POLYCLONAL ANTIBODYInjection day (e.g. day 0, 7, 30, etc.)AntigenAdjuvantAmount ant i g e n T o t a l i n j e c t i o n v o l u m e N u m b e r o f d i v i d e d i n j e c t i o n s I n j e c t i o n r o u t e a n d a n a t o m i c a l l o c a t i o n
( e . g . , S C , d o r s u m ) F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m g F O R M T E X T m L F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m g F O R M T E X T m L F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T F O R M T E X T m g F O R M T E X T m L F O R M T E X T F O R M T E X T
b . L i s t p o s s i b l e a d v e r s e e f f e c t s i n a n i m a l s t h a t m i g h t b e s e e n f r o m t h e p r o p o s e d a n t i g e n o r a d j u v a n t i n j e c t i o n s a n d w h a t m e a s u r e s w i l l b e t a k e n s h o u l d t h e s e a d v e r s e e f f e c t s o c c u r .
F O R M T E X T
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F O R M C H E C K B O X N o . P r o c e e d t o i t e m 4 .
F O R M C H E C K B O X Y e s . C o m p l e t e i t e m s 3 . c . i ) a n d i i ) ; t h e n p r o c e e d t o i t e m 4 .
i ) C o m p l e t e t h e f o l l o w i n g t a b l e . I n c l u d e a n y p r e - b l e e d s p r i o r t o i m m u n i z a t i o n s .
T A B L E 5 B L O O D C O L L E C T I O N S i t e o f b l o o d c o l l e c t i o n * A m o u n t o f b l o o d c o l l e c t e d e x p r e s s e d a s v o l u m e ( m L ) / a n d % o f b o d y w e i g h t ( a s s u m e 1 m l = 1 g r a m ) N u m b e r o f b l o o d c o l l e c t i o n s I n t e r v a l b e t w e e n c o l l e c t i o n s F O R M T E X T F O R M T E X T m L / F O R M T E X T % F O R M T E X T F O R M T E X T w e e k s F O R M T E X T F O R M T E X T m L / F O R M T E X T % F O R M T E X T F O R M T E X T w e e k s F O R M T E X T F O R M T E X T m L / F O R M T E X T % F O R M T E X T F O R M T E X T w e e k s
* T o a p p r o x i m a t e a c c e p t a b l e b l o o d c o l l e c t i o n v o l u m e s , a s s u m e t h e t o t a l b l o o d v o l u m e o f a h e a l t h y a n i m a l a s 7 % o f i t s b o d y w e i g h t , a n d t h a t 1 0 % o f t h e t o t a l b l o o d v o l u m e c a n b e c o l l e c t e d e v e r y 2 w e e k s w i t h o u t c a u s i n g s i g n i f i c a n t p h y s i o l o g i c a l c h a n g e . ( E x a m p l e : a c c e p t a b l e b l o o d v o l u m e f o r a 3 0 g m o u s e ( 0 . 0 7 X 3 0 g X 0 . 1 0 = . 2 1 m L . ) B l o o d v o l u m e s i n e x c e s s o f 1 0 % w i l l b e c o n s i d e r e d w i t h s c i e n t i f i c j u s t i f i c a t i o n . T y p i c a l l y t h e m a x i m u m b l o o d a v a i l a b l e f o r a t e r m i n a l p r o c e d u r e i s 5 0 % o f b l o o d v o l u m e - M U S T b e t a k e n w i t h t h e a n i m a l u n d e r a n e s t h e s i a .
i i ) W i l l a n e s t h e t i c s , t r a n q u i l i z e r s , o r a n a l g e s i c s b e u s e d p r i o r t o b l o o d c o l l e c t i o n ?
F O R M C H E C K B O X N o . J u s t i f y t h e o m i s s i o n o f p a i n - r e l i e v i n g a g e n t s b e l o w ; t h e n p r o c e e d t o i t e m 4 .
F O R M C H E C K B O X Y e s . D e s c r i b e t h e a d m i n i s t r a t i o n o f p a i n - r e l i e v i n g a g e n t s i n c l u d i n g d o s e ( m g / k g ) , v o l u m e ( m L ) , r o u t e , a n d f r e q u e n c y / d u r a t i o n b e l o w ; t h e n p r o c e e d t o i t e m 4 .
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4 . T e r m i n a l b l o o d c o l l e c t i o n : W i l l a n i m a l s u s e d f o r a n t i b o d y p r o d u c t i o n b e e x s a n g u i n a t e d a s a m e t h o d o f e u t h a n a s i a ?
F O R M C H E C K B O X N o . G o t o i t e m 5 .
F O R M C H E C K B O X Y e s . C o m p l e t e i t e m s 4 . a , b . , a n d c . b e l o w , t h e n g o t o i t e m 5 .
a . D e s c r i b e t h e m e t h o d o f e x s a n g u i n a t i o n .
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F O R M C H E C K B O X N o . J u s t i f y t h e o m i s s i o n o f p a i n - r e l i e v i n g a g e n t s b e l o w ; t h e n p r o c e e d t o i t e m 5 .
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VA Hospital @ # G Appendix AB Title
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_AuthorEmail _AuthorEmailDisplayName _ReviewingToolsShownOnce _dlc_DocId _dlc_DocIdItemGuid _dlc_DocIdUrl pu7 Replacement appendices JMorrison@ric.edu Morrison, Johanna 6 7 Z 3 Z X S P Z Z W Z - 5 0 9 - 4 % c e 5 c 6 1 0 6 - f e 5 8 - 4 3 3 c - b a f 2 - a d 3 5 2 8 a 7 f 5 7 d _ h t t p : / / w w w . r i c . e d u / i a c u c / _ l a y o u t s / 1 5 / D o c I d R e d i r . a s p x ? I D = 6 7 Z 3 Z X S P Z Z W Z - 5 0 9 - 4 , 6 7 Z 3 Z X S P Z Z W Z - 5 0 9 - 4