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4.²âÊÔ±¨¸æ£ºHPLC ͼÆ×£¬ Mass ͼÆ×£¬ COA ±¨¸æ
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N ¶ËÐÞÊÎ(Ac¡¢¹ó³¾´Ç³¦¡¢µþ´Ç³¦¡¢´Ü¡¢µþ³úµÈ)
N ¶ËÖ¬·¾ËáÐÞÊÎ(¶¹Þ¢Ëá¡¢×ØéµËá¡¢Ó²Ö¬Ëá µÈ)
C ¶ËÐÞÊÎ(-NH2¡¢-±Ê±·´¡¡¢-´¡²Ñ°ä¡¢-°¿²Ñ·¡¡¢-°¿·¡°ÕµÈ)
Ó«¹âÐÞÊÎ(FITC¡¢Biotin¡¢5-Fam¡¢ ÂÞµ¤Ã÷ µÈÔÚëĵÄN¶Ë£¬ëÄÁ´Öм䣬C¶ËÐÞÊÎ)
Á×ËữÐÞÊÎ(Thr(HPO3)¡¢Tyr(HPO3)¡¢Ser(HPO3) µÈ)
¼×»ù»¯ÐÞÊÎ(Lys(Me)¡¢Lys(Me)¡¢Lys(Me)¡¢Arg(Me) µÈ)
ÌØÊâ°±»ùËá(DÐÍ°±»ùËᣬbeta°±»ùËᣬhomo°±»ùËᣬ ·ÇÌìÈ»°±»ùËᣬ¸÷ÖÖ²àÁ´ÐÞÊÎ)
ÐÇ¿Õmv¹ú¾çÁº¼ÑоKLH¡¢BSA¡¢OVAżÁª·þÎñ (ͨ¹ý°ä²â²õ¡¢-±·±á2¡¢-°ä°¿°¿±áżÁª°³¢±á¡¢µþ³§´¡¡¢°¿³Õ´¡)
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·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо&²Ô²ú²õ±è; Àµ°±Ëá·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо (Lys¶þ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо¡¢³¢²â²õËÄ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо¡¢³¢²â²õ°Ë·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо)
Äñ°±Ëá·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо (Orn¶þ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо¡¢°¿°ù²ÔËÄ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо¡¢°¿°ù²Ô°Ë·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо)
²»¶Ô³Æ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо&²Ô²ú²õ±è; Lys²»¶Ô³Æ¶þ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо£¬&²Ô²ú²õ±è;°¿°ù²Ô²»¶Ô³Æ¶þ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо
Glu²àÁ´ÐÇ¿Õmv¹ú¾çÁº¼Ñо£¬´¡²õ±è²àÁ´ÐÇ¿Õmv¹ú¾çÁº¼Ñо&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;&²Ô²ú²õ±è;
»·×´ÐÇ¿Õmv¹ú¾çÁº¼Ñо&²Ô²ú²õ±è; ͨ¹ý¶þÁò¼ü³É»·µÄÐÇ¿Õmv¹ú¾çÁº¼Ñо&²Ô²ú²õ±è;
µ¥¶þÁò¼üÐÇ¿Õmv¹ú¾çÁº¼Ñо£¬Ë«¶þÁò¼üÐÇ¿Õmv¹ú¾çÁº¼Ñо£¬Èþ¶Ô¶þÁò¼üÐÇ¿Õmv¹ú¾çÁº¼Ñо£¬
·Ö×Ó¼ä¶Ô³Æ¶þÁò¼üÐÇ¿Õmv¹ú¾çÁº¼Ñо£¬·Ö×Ӽ䲻¶Ô³Æ¶þÁò¼üÐÇ¿Õmv¹ú¾çÁº¼Ñо
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7. ²»Í¬¹æÄ££º
µ¥ÌõÐÇ¿Õmv¹ú¾çÁº¼Ñо ÅúÁ¿ÐÇ¿Õmv¹ú¾çÁº¼Ñо
ëÄ¿â&²Ô²ú²õ±è; [ÖصþëÄ¿â, °±»ùËáɸѡëÄ¿â, ¶¨Î»É¸Ñ¡ëÄ¿â, Ëæ»úëÄ¿â(ÔÓÂÒëÄ¿â),
½Ø¶ÌëĿ⣬µ¥Î»µãɨÃèëÄ¿â&²Ô²ú²õ±è; ˫λµã×éºÏɨÃèëÄ¿â Èýλµã×éºÏɨÃèëÄ¿â]
ÐÇ¿Õmv¹ú¾çÁº¼Ñо¶¨Öƺϳɰ¸Àýչʾ
1. ÏßÐÔÐÇ¿Õmv¹ú¾çÁº¼Ñо&²Ô²ú²õ±è;
a. H-Arg-Gly-Asp-Cys-OH
b. DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA
2. ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо
a. -Leu-beta-Ala
b. [Orn(SDGGDADS)2]-Leu-beta-Ala
3. ²»¶Ô³Æ·ÖÖ§ÐÇ¿Õmv¹ú¾çÁº¼Ñо
a. Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Leu-Glu-Gly-Gln-Ala-Ala-Lys(gama-Glu-Pal)-Glu-
Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH
b. SDGGDADSK(SDSDG)-WDGDS
4. »·×´ÐÇ¿Õmv¹ú¾çÁº¼Ñо
a. D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-Ol (Disulfide bridge Cys2-Cys7)
b. Ile-Lys-Cys-Asn-Cys-Lys-Arg-His-Val-Ile-Lys-Pro-His-Ile-Cys-Arg-Lys-Ile-Cys-Gly-Lys-Asn-NH2 (Disulfide bridge Cys3-Cys15, Cys5-Cys19)
5. N ¶ËÐÞÊÎ&²Ô²ú²õ±è;(Ac¡¢¹ó³¾´Ç³¦¡¢´Ü¡¢µþ³úµÈ)
a. Fmoc-Lys-Ala-Ala
b. Ac-Lys-Ala-Ala-Lys-Leu-NH2
6. N¶ËÖ¬·¾ËáÐÞÊÎ(¶¹Þ¢Ëá¡¢×ØéµËá¡¢Ó²Ö¬Ëá µÈ)
a.Myr-KKKKKSGKSE
b.Pal-KKKKKSGKSE
7. C ¶ËÐÞÊÎ(-NH2¡¢-±Ê±·´¡¡¢-´¡²Ñ°ä¡¢-°¿²Ñ·¡¡¢-°¿·¡°ÕµÈ)
a. Boc--Leu-Gly-Arg-PNA.HCL
b. Boc--Leu-Gly-Arg-AMC
8. Ó«¹âÐÞÊÎ(FITC¡¢Biotin¡¢5-Fam¡¢ ÂÞµ¤Ã÷ µÈÔÚëĵÄN¶Ë£¬ëÄÁ´Öм䣬C¶ËÐÞÊÎ)
a. FITC-Lys-Cys-Asn-Cys-Lys-Arg-His-Val-Ile-Lys-Pro-His-Ile-Cys-Arg-Lys
b. Lys-Cys-Asn-Cys-Lys(Biotin)-Arg-His-Val-Ile-Lys-Pro-His-Ile-Cys-Arg-Lys.
9. Á×ËữÐÞÊÎ (Thr(HPO3)¡¢Tyr(HPO3)¡¢Ser(HPO3) µÈ)
a. Ac-Ile-Tyr(PO3H2)-Gly-Glu-Phe-NH2
b. Phe-Phe-Lys-Asn-Ile-Val-Thr-Pro-Arg-Thr(PO3H2)-Pro-Pro-Pro-Ser-Gln-Gly-Lys-NH2
10. ¼×»ù»¯ÐÞÊÎ(Lys(Me)¡¢Lys(Me)¡¢Lys(Me)¡¢Arg(Me) µÈ)
a. Asn-Cys-Lys-Ile-Lys(me)-Cys-Arg-His-Val-Ile-Lys-Pro
b. Cys-Lys-Arg-His-Ile-Lys(me)3-Cys-Asn-Val-Ile-Lys-Pro
c. Arg-His-Val-Ile-Ile-Lys-Cys-Asn-Cys-Lys-Arg(Me)-Pro
11. ÌØÊâ°±»ùËá(DÐÍ°±»ùËá¡¢²ú±ð³Ù²¹°±»ùËá¡¢³ó´Ç³¾´Ç°±»ùËá¡¢·ÇÌìÈ»°±»ùËá¡¢²àÁ´´øÓÐÌØÊâÐÞÊÎ)
a. D-Arg-Arg-Pro-Hyp-Gly-Thi-Ser-D-Tic-Oic
b. c[Mpr-D-Tyr(OEt)-Ile-Thr-Asn-Cys]-Pro-Orn-Gly-NH2
c. Ac-3-(2-naphthyl)-D-Ala-4-Chloro-D-Phe-3(3-pyridyl)-D-Ala-D-Cit-Leu-Arg-Pro-D-Ala-NH2
12. ¸´ÔÓÐÇ¿Õmv¹ú¾çÁº¼Ñо
a. Biotin-Ile-Lys(me)-Cys-Asn-Cys-Lys-Arg-His-Val-Ile-Lys-Pro-Thr(HPO3)-Leu-NH2
b. E[c(RGDfK)]2
13. ëĿ⣺
ÖصþëÄ¿â, °±»ùËáɸѡëÄ¿â¡¢¶¨Î»É¸Ñ¡ëÄ¿â¡¢Ëæ»úëÄ¿â(ÔÓÂÒëÄ¿â),
½Ø¶ÌëÄ¿â¡¢µ¥Î»µãɨÃèëĿ⡢˫λµã×éºÏɨÃèëÄ¿â¡¢Èýλµã×éºÏɨÃèëÄ¿â&²Ô²ú²õ±è;
14. ÐÇ¿Õmv¹ú¾çÁº¼Ñо°³¢±á¡¢µþ³§´¡¡¢°¿³Õ´¡Å¼Áª·þÎñ&²Ô²ú²õ±è;&²Ô²ú²õ±è;£º
Cys-Asn-Lys-Ile-Lys-Arg--Arg-His-Val-Ile-Lys-Pro ͨ¹ý°ä²â²õżÁª°³¢±á
Asn-Lys-Ile-Lys-Arg-Arg-His-Val-Ile-Lys-Pro ͨ¹ýôÈ»ùżÁªµþ³§´¡
ÐÇ¿Õmv¹ú¾çÁº¼ÑоÐÞÊαí
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥±·¶ËÐÞÊÎ | ||||||||||||
Ac | Fmoc | CBZ | Boc | Bz | Mpa- | Tos | For | |||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥±·¶ËÁ¬½ÓÖ¬·¾Ëá | ||||||||||||
Myr- | Suc | MeOSuc | Ste- | Hex- | Oct- | Dec- | Pal- | |||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥°ä¶ËÐÞÊÎ | ¡¡ | ¡¡ | ¡¡ | ¡¡ | ||||||||
-pNA | -OMe | -ol | -OBzl | -NH2 | -OEt | -AMC | -NHEt | |||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥¶Ù¹¹ÐÍ°±»ùËá | ||||||||||||
D-Ala | D-Lys | D-Glu | D-Tyr | D-Asp | D-Pro | D-His | D-Trp | |||||
D-Arg | D-Met | D-Gln | D-Thr | D-Asn | D-Phe | D-Leu | D-Val | |||||
D-Cys | D-Ser | ¡¡ | ¡¡ | ¡¡ | ¡¡ | ¡¡ | ¡¡ | |||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥·ÇÌìÈ»°±»ùËá | ||||||||||||
Beta-Asp | 3-Thi | Chg | Cha | D-1-Nal | Met(O) | Nle | Pip | |||||
D-Beta-Asp | Aib | D-Chg | D-Cha | L-1-Nal | D-Met(O) | D-Nle | Nip | |||||
Gamma-Glu | Abu | Dab | Oic | D-2-Nal | Cys(SNO) | Cit | 2-Aze | |||||
D-Gamma-Glu | D-Abu | Dap | Tic | L-2-Nal | Met(O)2 | D-Cit | 3-Aze | |||||
D-4-Pal | Hyp | Pra | Chg | D-2-Pal | D-Met(O)2 | Orn | Se-Cys | |||||
L-4-Pal | Phg | D-Pra | Cys(Bzl) | L-2-Pal | Lys(Ac) | D-Orn | DOPA | |||||
D-3-Pal | D-Phg | Allo-Thr | Epsilon-Lys | isoGln | Ac-Lys | Pen | ¡¡ | |||||
L-3-Pal | Nva | D-Allo-Thr | D-Epsilon-Lys | D-isoGln | Lys(Dde) | D-Pen | ¡¡ | |||||
Tle | D-Nva |
|
| ¡¡ | ¡¡ | ¡¡ | ¡¡ | |||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥Ó«¹âÐÇ¿Õmv¹ú¾çÁº¼Ñо | ||||||||||||
Biotin- | FITC- | 5-FAM- | Lys(Dnp) | |||||||||
Biotin-Ahx- | FITC-Ahx- | 5-FAM-Ahx- | D-Lys(Dnp) | |||||||||
-Lys(Biotin)- | -Lys(FITC)- | -Lys(5-FAM)- | Dab(Dnp) | |||||||||
-Lys(Biotin) | -Lys(FITC) | Lys(5-FAM) | Dap(Dnp) | |||||||||
5-TAMRA- | -Lys(TAMRA)- | ¡¡-³¢²â²õ(5-°Õ´¡²Ñ¸é´¡) | ¡¡ | |||||||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥°¿³Õ´¡,°³¢±á,µþ³§´¡µ°°×żÁª | ||||||||||||
BSA-Peptide N terminus | KLH-Peptide N terminus | OVA-Peptide N terminus | ||||||||||
BSA-peptide C terminus | KLH-Peptide C terminus | OVA-peptide C terminus¡¡ | ||||||||||
BSA-Peptide via Cys | KLH-peptide via Cys | OVA-peptide via Cys¡¡ | ||||||||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥¼×»ù»¯ÐÞÊÎ | ||||||||||||
Lys(Me) | Arg(Me) | Tyr(Me) | Tyr(Et) | |||||||||
Lys(Me2) | Arg(Me)2 asymmetrical | Thr(Me) | D-Tyr(Et) | |||||||||
Lys(Me3) | Arg(Me)2 symmetrical | Ser(Me) | ¡¡ | |||||||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥Á×ËữÐÇ¿Õmv¹ú¾çÁº¼Ñо | ||||||||||||
pSer | pTyr | pThr | Di-sites in sequence | |||||||||
D-pSer | D-pTyr | D-pThr | Tri-sites in sequence | |||||||||
ÐÇ¿Õmv¹ú¾çÁº¼ÑоºÏ³É´¥ëÄÊ÷Ö¬ | ||||||||||||
peptide on Wang Resin | peptide on 2-CTC Resin | |||||||||||
Peptide on Rink Amide AM Resin | peptide on Rink Amide MBHA Resin | |||||||||||
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