PMID- 17027283
OWN - NLM
STAT- MEDLINE
DCOM- 20070509
LR  - 20171116
IS  - 1046-5928 (Print)
IS  - 1046-5928 (Linking)
VI  - 52
IP  - 1
DP  - 2007 Mar
TI  - Preparation and characterization of recombinant protein ScFv(CD11c)-TRP2 for 
      tumor therapy from inclusion bodies in Escherichia coli.
PG  - 131-8
AB  - Dendritic cells (DCs)-based immunotherapy represents an approach to the 
      prevention and treatment of cancers. Targeting antigens to receptors on DCs can 
      be expected to enhance immune response. We have constructed an expression vector 
      pET32a(+)-ScFv(CD11c)-TRP2 based on a single-chain antibody fragment (ScFv) that 
      targets the high affinity receptor CD11c which is expressed on murine DCs. The 
      3'-terminal end of the ScFv was ligated to the gene for MHC class I 
      molecule-recognized peptide from mouse tyrosine-related protein 2 (TRP2). Using 
      this vector, we have expressed and purified ScFv(CD11c)-TRP2, a fusion protein 
      that could target TRP2 peptide to CD11c on DCs in vivo to elicit anti-tumor 
      responses. This fusion protein was expressed in inclusion bodies in Escherichia 
      coli BL21(DE3) and was refolded and purified on-column effectively by immobilized 
      metal affinity chromatography using His-tag. Flow cytometry assays showed the 
      specific binding ability of ScFv(CD11c)-TRP2 to DCs, which could be blocked by a 
      hamster anti-mouse CD11c produced by N418 hybridoma. Further studies demonstrated 
      that ScFv(CD11c)-targeted TRP2 peptide processed by DCs was capable of 
      stimulating T cells proliferation. Thus, this fusion protein provides a basis for 
      further research in cancer therapy in vivo.
FAU - Kou, Geng
AU  - Kou G
AD  - International Joint Cancer Institute, Second Military Medical University, New 
      Library Building West 10th-11th Floor, 800 Xiang Yin Road, Shanghai 200433, PR 
      China.
FAU - Shi, Shu
AU  - Shi S
FAU - Wang, Hao
AU  - Wang H
FAU - Tan, Min
AU  - Tan M
FAU - Xue, Jingya
AU  - Xue J
FAU - Zhang, Dapeng
AU  - Zhang D
FAU - Hou, Sheng
AU  - Hou S
FAU - Qian, Weizhu
AU  - Qian W
FAU - Wang, Shuhui
AU  - Wang S
FAU - Dai, Jianxin
AU  - Dai J
FAU - Li, Bohua
AU  - Li B
FAU - Guo, Yajun
AU  - Guo Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060823
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (Antigens, CD)
RN  - 0 (Antineoplastic Agents)
RN  - 0 (CD11c Antigen)
RN  - 0 (DNA Primers)
RN  - 0 (Immunoglobulin Variable Region)
RN  - 0 (Membrane Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (peptide SVYDFFVWL)
SB  - IM
MH  - Animals
MH  - Antigens, CD/genetics/therapeutic use
MH  - Antineoplastic Agents/therapeutic use
MH  - Base Sequence
MH  - CD11c Antigen/genetics/therapeutic use
MH  - Cloning, Molecular
MH  - DNA Primers
MH  - Dendritic Cells/drug effects/immunology
MH  - Escherichia coli/genetics
MH  - Humans
MH  - Immunoglobulin Variable Region/chemistry/*genetics/isolation & 
      purification/therapeutic use
MH  - Membrane Proteins/chemistry/genetics
MH  - Mice
MH  - Neoplasms/*drug therapy
MH  - Peptide Fragments/chemistry/genetics
MH  - Protein Folding
MH  - Recombinant Proteins/chemistry/isolation & purification/therapeutic use
MH  - Restriction Mapping
EDAT- 2006/10/10 09:00
MHDA- 2007/05/10 09:00
CRDT- 2006/10/10 09:00
PHST- 2006/07/25 00:00 [received]
PHST- 2006/08/11 00:00 [accepted]
PHST- 2006/10/10 09:00 [pubmed]
PHST- 2007/05/10 09:00 [medline]
PHST- 2006/10/10 09:00 [entrez]
AID - S1046-5928(06)00256-7 [pii]
AID - 10.1016/j.pep.2006.08.007 [doi]
PST - ppublish
SO  - Protein Expr Purif. 2007 Mar;52(1):131-8. doi: 10.1016/j.pep.2006.08.007. Epub 
      2006 Aug 23.