Part:BBa_K3090000
cell penetrating peptide
nuclear localization signal (NLS) enriched in positively charged residues that can function as a Cell-penetrating peptide (CPP), similar to the classical CPP derived from HIV type 1 transactivator of transcription protein (HIV TAT).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
This part(BBa_K3090000) was fused to the N-terminus of scFv(P5) (part number BBa_K3090001) to design a cell-penetrating antibody fragment (part BBa_K3090002). The sequence of the designed protein is as follows.
MTYTRRRFRR RRHRPRS QVQLQESGGD LVQPGGSLKL SCAVSGFSLT GYGVNWVRQT PDKRLEWVAM IWGDGNTDYN SSVKGRFTIS KDNAKSTVYL QMSSLKSEDT AMYYCARERD YRLDYWGQGT TVTVSS GGGGSGGGGSGGGGS DIELTQSPAS LAVSLGQRAT ISCRASGNIH NYLAWYQQKP GQPPKLLIYY TTTLADGIPA RFSGSGSGTD YTLTINPVEA DDVATYYCQH FWSTPRTFGG GTKLEIKR
In 2019, a paper is published about the efficiency of fusion peptides for intracellular delivery of therapeutic antibodies.
<Reference> Gaston, J., Maestrali, N., Lalle, G., Gagnaire, M., Masiero, A., Dumas, B., Dabdoubi, T., Radošević, K., Berne, P.-F. Intracellular delivery of therapeutic antibodies into specific cells using antibody-peptide fusions (2019) Scientific Reports, 9 (1), art. no. 18688,
In this paper, the authors investigated various fusion peptides for their efficiency.
Name CPP_length Properties Net_charge CPP_Sequence
Pep-1 21 Amphipathic 3 KETWWETWWTEWSQPKKKRKV
TAT 11 Cationic 8 YGRKKRRQRRR
PEPth 12 Cationic 5 VKKKKIKAEIKI
aurein 1.2 13 Amphipathic 1 GLFDIIKKIAESF
MTS 17 Hydrophobic 0 KGEGAAVLLPVLLAAPG
GFWFG 5 Hydrophobic 0 GFWFG 31
Above peptides are attached to different positions of antibody (N-terminal of light or heavy chain, C-terminal of light or heavy chain, hinge region of heavy chain) and the antibody yield and efficiency of delivery were investigated. The result showed Pep-1 and PEPth were best when attached to C-terminal of light chain or hinge region.
We believe this study can be useful for teans who wants to design a fusion peptide to deliver an anitbody into the cell.
References
1: Yu W, Zhan Y, Xue B, Dong Y, Wang Y, Jiang P, Wang A, Sun Y, Yang Y. Highly efficient cellular uptake of a cell-penetrating peptide (CPP) derived from the capsid protein of porcine circovirus type 2. J Biol Chem. 2018 Sep 28;293(39):15221-15232.
2. Gaston, J., Maestrali, N., Lalle, G., Gagnaire, M., Masiero, A., Dumas, B., Dabdoubi, T., Radošević, K., Berne, P.-F. Intracellular delivery of therapeutic antibodies into specific cells using antibody-peptide fusions (2019) Scientific Reports, 9 (1), art. no. 18688
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