Construction and expression of CD3/GD2 BiTE.

(A) Schematic diagram of CD3/GD2 BiTE in pcDNA3.1(+). CD3/GD2 BiTE consists of human scFv-CD3 clone 18 (VH-VL) linked to mouse scFv-GD2 (VL-VH) via a (G4S)3 linker. An Interleukin-2 signal peptide for protein secretion and a polyhistidine (His6) tag...

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محفوظ في:

التفاصيل البيبلوغرافية
المؤلف الرئيسي:	Kuntida Kitidee (3935783) (author)
مؤلفون آخرون:	Sumet Amonyingcharoen (21512526) (author), Sarinthip Preedagasamzin (15332756) (author), Korakot Atjanasuppat (3821833) (author), Piamsiri Sawaisorn (14608498) (author), Pornprapa Srimorkun (21512529) (author), Sawang Petvises (523491) (author), Wanpen Chaicumpa (17781) (author), Suparerk Borwornpinyo (3509102) (author), Usanarat Anurathapan (4570963) (author), Suradej Hongeng (285857) (author)
منشور في:	2025
الموضوعات:	Biophysics Biochemistry Cell Biology Molecular Biology Neuroscience Physiology Pharmacology Biotechnology Immunology Developmental Biology Cancer Hematology Infectious Diseases Virology Space Science Biological Sciences not elsewhere classified indirect elisa assay flexible peptide linker div >< p compact monomeric structure 2 signal sequence treating various types gd bite armed cells could represent alternative immunotherapy strategy alternative immunotherapy method bind cd3 epsilon treating neuroblastoma patients gd2 bite combined positive jurkat cells negative raji cells mouse scfv gd2 vh ), linked neuroblastoma cell lines dimensional cell cultures combining bispecific antibody treating neuroblastoma gd2 bite negative sk nb ), human scfv combining cd3 bispecific antibody neuroblastoma cells sy5y cells cells enabled cell engager gd2 bispecific positive sh vitro </ transiently produced target antigens sy5y gfp strong potential results suggest protein secretion promising approach polyhistidine tag intriguing targets human vγ9vδ2 highest ability high yield cytotoxic abilities binding specificity based strategies 360 nm
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الوصف
الملخص:	<p>(A) Schematic diagram of CD3/GD2 BiTE in pcDNA3.1(+). CD3/GD2 BiTE consists of human scFv-CD3 clone 18 (VH-VL) linked to mouse scFv-GD2 (VL-VH) <i>via</i> a (G<sub>4</sub>S)3 linker. An Interleukin-2 signal peptide for protein secretion and a polyhistidine (His6) tag for protein purification and detection were added to the N-terminus and C-terminus of CD3/GD2 BiTE, respectively. A simplified structural model of CD3/GD2 BiTE is presented below. (B) Amino acid sequence of CD3/GD2 BiTE. The CDRs in the VH and VL regions of human scFv CD3 clone 18 and mouse scFv GD2 are indicated by colored and underlined text. The linkers and polyhistidine (His6) tag are highlighted in green. (C) Reducing-PAGE analysis of purified CD3/GD2 BiTE protein stained with InstantBlue and western blot analysis of purified CD3/GD2 BiTE detected using an anti-histidine tag antibody. The molecular weight of CD3/GD2 BiTEs is approximately 58 kDa. (D) Non-reducing-PAGE analysis of CD3/GD2 BiTE treated with DTT at different concentrations, stained with InstantBlue<sup>TM</sup>. CD3/GD2 BiTE was dissociated by DTT and migrated on non-reducing-PAGE.</p>

Construction and expression of CD3/GD2 BiTE.

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