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1 | RAMS-2.3.0.zip\RAMS-2.3.0\Webapp\RAMS-2.3.0\RAMS-2.3.0.war\WEB-INF\classes | ACORP_Appendix2_59.docx | Wed May 2 23:25:58 2018 UTC |
2 | RAMS-2.3.0.zip\RAMS-2.3.0\Webapp\RAMS-2.3.0\RAMS-2.3.0.war\WEB-INF\classes | ACORP_Appendix2_59.docx | Tue Jul 17 21:28:04 2018 UTC |
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1 | Official D ate of App roval► | |
2 | ||
3 | ACORP Appe ndix 2 | |
4 | Antibody P roduction | |
5 | Version 4 | |
6 | ||
7 | See ACORP App. 2 Ins tructions, for more detailed e xplanation s of the i nformation requested . | |
8 | ||
9 | Immunizati on. Provi de the inf ormation r equested b elow for a ny animals to be use d for rais ing antibo dies speci fically fo r use in t his protoc ol. | |
10 | ||
11 | Describe t he immuniz ation prot ocol in th e table be low, using a separat e row for each day o n which an y agent (i ncluding p rimer, ant igen, and/ or adjuvan t) will be administe red. (Mak e sure tha t each pri mer, antig en, and ad juvant is also inclu ded in App endix 3.) | |
12 | ||
13 | Immun-izat ion day | |
14 | (e.g. day -7, 0, 7, 30, etc.) | |
15 | Antigen | |
16 | Adjuvant – give name , concentr ation, and volume (m l) | |
17 | Total inje ction volu me (ml) pe r animal ( antigen pl us adjuvan t) | |
18 | Divided am ong how ma ny injecti on sites? | |
19 | Injection route and location o f injectio n site(s) on body | |
20 | ||
21 | Name | |
22 | Total amou nt (mg) an d volume ( ml) | |
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48 | ||
49 | Describe h ow each an tigen will be screen ed to make sure that it does n ot harbor infectious agents th at could i nfect othe r laborato ry animals or people after inj ection. | |
50 | ► | |
51 | ||
52 | List possi ble advers e effects that might be observ ed in anim als receiv ing the pr oposed pri mer, antig en, and/or adjuvant injections , and desc ribe the m easures th at will be taken if these adve rse effect s occur: | |
53 | ► | |
54 | ||
55 | Give the j ustificati on for usi ng any pri mer or adj uvant that is expect ed to caus e pain or distress i n the anim als. | |
56 | ► | |
57 | ||
58 | Survival B lood Colle ction. Wi ll blood b e collecte d as a sur vival proc edure for the produc tion and h arvesting of antibod ies on thi s protocol ? | |
59 | ||
60 | ► ( ) No , the prod uction and harvest o f antibodi es on this protocol does not i nvolve sur vival coll ection of blood. | |
61 | ||
62 | ► ( ) Ye s, this pr otocol req uires the collection of blood in a survi val proced ure, befor e (as a “p re-bleed”) and/or af ter immuni zation. M ake sure t his is inc luded in I tem R of t he ACORP, and comple te items 2 .a, 2.b, a nd 2.c, be low. | |
63 | ||
64 | Describe e ach surviv al collect ion of blo od in the table belo w, includi ng any “pr e-bleeds” prior to i mmunizatio ns: | |
65 | ||
66 | Site of Bl ood Collec tion | |
67 | Amount of Blood Coll ected at a ny one tim e, | |
68 | expressed as volume (ml) and a s % of bod y weight ( assume 1 m l = 1 gram ) | |
69 | Number of Blood Coll ections | |
70 | Time Inter val(s) Bet ween Succe ssive Coll ections | |
71 | Volume Rep lace-ment? | |
72 | (yes/no) | |
73 | ||
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88 | ||
89 | Will anest hetics, tr anquilizer s, or anal gesics be administer ed for blo od collect ion? | |
90 | ||
91 | ► ( ) No anesthetic s, tranqui lizers, or analgesic s will be administer ed for blo od collect ion. Expl ain why it is approp riate or n ecessary N OT to admi nister pai n-relievin g agents: | |
92 | ► | |
93 | ||
94 | ► ( ) Yes . Describ e the admi nistration of pain-r elieving a gents, inc luding the name of e ach agent, and its d ose (mg/kg ), volume (ml), and route and frequency/ duration o f administ ration (Ma ke sure th is informa tion is al so include d in Appen dix 3): | |
95 | ► | |
96 | ||
97 | Will volum e replacem ent be pro vided for blood that is collec ted? | |
98 | ||
99 | ► ( ) Vol ume will N OT be repl aced for s ome of the blood col lection li sted. For each coll ection lis ted in Ite m 2.a, abo ve, for wh ich volume will NOT be replace d, explain why not. | |
100 | ► | |
101 | ||
102 | ► ( ) Vol ume WILL b e replaced for some of the blo od collect ion listed . For eac h collecti on listed in Item 2. a, above, for which volume WIL L be repla ced, descr ibe the re placement( s) that wi ll be prov ided (incl uding the compositio n of the r eplacement (s), volum e, and rou te of admi nistration ). | |
103 | ► | |
104 | ||
105 | Terminal B lood Colle ction. Wi ll animals be euthan atized by exsanguina tion, for harvest of antibodie s? | |
106 | ||
107 | ► ( ) No , this pro tocol does NOT invol ve termina l blood co llection f or harvest of antibo dies. | |
108 | ► ( ) Ye s, this pr otocol DOE S require terminal b lood colle ction for the harves t of antib odies. Ma ke sure th is is incl uded in It em R of th e ACORP, a nd complet e Items 3. a., 3. b., and 3.c., below: | |
109 | ||
110 | Describe t he method( s) to be u sed for eu thanasia a nd exsangu ination: | |
111 | ► | |
112 | ||
113 | Will anest hetics, tr anquilizer s, or anal gesics be administer ed for exs anguinatio n? | |
114 | ||
115 | ► ( ) No anestheti cs, tranqu ilizers, o r analgesi cs will be administe red for th e exsangui nation(s). Explain why it is appropriat e or neces sary NOT t o administ er pain-re lieving ag ents: | |
116 | ► | |
117 | ||
118 | ► ( ) Yes . Describ e the admi nistration of pain-r elieving a gents incl uding the name of ea ch agent, and its do se (mg/kg) , volume ( ml), and r oute and f requency/d uration of administr ation (Mak e sure thi s informat ion is als o included in Append ix 3): | |
119 | ► | |
120 | ||
121 | Describe h ow you wil l make sur e that the animals a re dead af ter collec tion of th e blood: | |
122 | ► | |
123 | ||
124 | Harvesting Feeder Ce lls. Desc ribe the e xact proce dures (inc luding adm inistratio n of pain- relieving agents) th at will be used on a ny donor a nimals fro m which fe eder cells will be c ollected f or this pr otocol, an d estimate the numbe r of anima ls needed for this p urpose. Ma ke sure th at these a nimals are included in Item I of the ACO RP, and th at the har vesting of feeder ce lls is inc luded in I tem R of t he ACORP. | |
125 | ► | |
126 | ||
127 | Expansion of Hybrido ma Cell Li ne(s) in v ivo. Will any anima ls be used to expand hybridoma cell line s so that antibody c an be harv ested from ascites f luid? | |
128 | ||
129 | ► ( ) No animals w ill be use d on this protocol f or in vivo expansion of hybrid oma cell l ines. | |
130 | ► ( ) Ye s, this pr otocol req uires use of some an imals for in vivo ex pansion of hybridoma cell line s. Make s ure that t he animals used for this are i ncluded in Item I of the ACORP , the prim ing agent and the hy bridoma ce lls are do cumented i n Appendix 3, and th e collecti on of asci tes fluid is include d in Item R of the A CORP. Com plete item s 5.a, 5.b , and 5.c, below. | |
131 | ||
132 | Explain wh y alternat e research methods t hat do not require t he use of additional animals ( e.g., in v itro cell culture sy stems for harvesting monoclona l antibodi es) are no t adequate to meet t he researc h objectiv es of this project. | |
133 | ► | |
134 | ||
135 | Complete t he followi ng table t o summariz e the proc edures to be perform ed in expa nding the hybridoma cell lines and colle cting asci tes fluid: | |
136 | ||
137 | Hybridoma cell line designatio n | |
138 | Number of animals to be used f or ascites productio n | |
139 | Priming ag ent and vo lume | |
140 | Number and timing of priming i njections | |
141 | Volume of injected h ybridoma c ells | |
142 | Number of abdominal taps befor e euthanas ia | |
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161 | ||
162 | Describe t he exact p rocedures (including administr ation of p ain-reliev ing agents ) that wil l be used for the ab dominal ta ps to be p erformed o n this pr otocol | |
163 | ► | |
164 | ||
165 | List the c riteria fo r euthanas ia of anim als prior to the las t planned abdominal tap. | |
166 | ► | |
167 | ||
168 | (Use Appen dix 9 to d ocument an y “departu res” from the standa rds in the Guide rep resented b y these pr ocedures. Consult th e IACUC or the Atten ding Veter inarian fo r help in determinin g whether any “depar tures” are involved. ) |
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