A phage library-derived single-chain Fv fragment in complex with turkey egg-white lysozyme: characterization, crystallization and preliminary X-ray analysis

1998 ◽  
Vol 35 (3) ◽  
pp. 189-194 ◽  
Author(s):  
G Küttner ◽  
T Keitel ◽  
E Gieimann ◽  
H Wessner ◽  
C Scholz ◽  
...  
Keyword(s):  
Egg White ◽  
Phage Library ◽  
Single Chain ◽  
X Ray ◽  
Egg White Lysozyme ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

Preliminary X-ray investigation of duck egg-white lysozyme II

1972 ◽  
Vol 71 (3) ◽  
pp. 815-817 ◽  
Author(s):  
J. Berthou ◽  
A. Laurent ◽  
P. Jolle`s
Keyword(s):  
Egg White ◽  
X Ray ◽  
Egg White Lysozyme ◽  
Duck Egg

Anti-Human Epidermal Growth Factor Receptor 2 Single-Chain Fv Fragment-Decorated DM1 Nanoparticles for Specific Targeting of Human Epidermal Growth Factor Receptor 2-Positive Breast Tumor Cells

2021 ◽  
Vol 17 (3) ◽  
pp. 447-455
Author(s):  
Ling Ni ◽  
You-Xin Li
Keyword(s):  
Breast Cancer ◽  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Growth Factor Receptor ◽  
Single Chain ◽  
Human Epidermal Growth Factor ◽  
Single Chain Fv ◽  
Epidermal Growth ◽  
Single Chain Fv Fragment

Purpose: Although monoclonal antibodies are used to decorate nanoparticles to target specific cells, penetration of tumor tissues by monoclonal antibodies is limited by their large size. Therefore, we prepared DM1 nanoparticles decorated with the small anti-HER2 single-chain Fv fragment (scFvHER2) of trastuzumab (TMAB) for targeting to human epidermal growth factor receptor 2 (HER2) overexpressing in breast cancer effectively. Methods: ScFvHER2 fragment was coupled with DM1 nanoparticles (NPs) via covalent thiol-maleimide linkages. Their physicochemical properties, uptake by cells, and toxicity to tumor cells were investigated. Their vivo biodistribution was assessed employing liquid chromatographytandem mass spectrometry, while their antitumor activity was investigated in nude mice burdened with BT-474 tumor. Results: Viability of BT-474 cells incubated with scFvHER2-DM1-Nanoparticles (scFv-DM1-NPs) was significantly lower than that of BT-474 cell treated with TMAB-DM1-Nanoparticles (TMAB-DM1-NPs) (P < 0 05). Uptake by cells of scFvDM1-NPs was significantly higher than TMAB-DM1-NPs (P < 0 01). Accumulation of scFv-DM1-NPs in tumor tissue was notably higher than TMAB-DM1-NPs (P < 0 05). scFv-DM1-NPs exhibited improved antitumor effects compared to TMABDM1-NPs (P < 0 05), showing a tumor inhibition rate of more than 70%. Conclusions: ScFvHER2 fragment could serve as a more effective targeting ligand than TMAB, and scFv-DM1-NPs could be developed as a possible drug delivery system to target HER2-positive breast cancer.

Analysis of polycrystallinity in hen egg-white lysozyme using a pnCCD

2012 ◽  
Vol 45 (3) ◽  
pp. 517-522 ◽  
Author(s):  
Sebastian Send ◽  
Ali Abboud ◽  
Wolfram Leitenberger ◽  
Manfred S. Weiss ◽  
Robert Hartmann ◽  
...  
Keyword(s):  
Sample Surface ◽  
Egg White ◽  
Hen Egg White Lysozyme ◽  
X Ray ◽  
Lattice Constants ◽  
Egg White Lysozyme ◽  
Hen Egg White ◽  
Diffraction Patterns ◽  
Crystalline Domains ◽  
Hen Egg

A crystal of hen egg-white lysozyme was analyzed by means of energy-dispersive X-ray Laue diffraction with white synchrotron radiation at 2.7 Å resolution using a pnCCD detector. From Laue spots measured in a single exposure of the arbitrarily oriented crystal, the lattice constants of the tetragonal unit cell could be extracted with an accuracy of about 2.5%. Scanning across the sample surface, Laue images with split reflections were recorded at various positions. The corresponding diffraction patterns were generated by two crystalline domains with a tilt of about 1° relative to each other. The obtained results demonstrate the potential of the pnCCD for fast X-ray screening of crystals of macromolecules or proteins prior to conventional X-ray structure analysis. The described experiment can be automatized to quantitatively characterize imperfect single crystals or polycrystals.

Crystal distortion of monoclinic hen egg-white lysozyme crystals using X-ray digital topography

2014 ◽  
Vol 401 ◽  
pp. 238-241
Author(s):  
Kei Wako ◽  
Daiki Fujii ◽  
Shiro Tsukashima ◽  
Takeharu Kishi ◽  
Masaru Tachibana ◽  
...  
Keyword(s):  
Egg White ◽  
Hen Egg White Lysozyme ◽  
X Ray ◽  
Egg White Lysozyme ◽  
Hen Egg White ◽  
Lysozyme Crystals ◽  
Hen Egg

Digital topography with an X-ray CCD camera for characterizing perfection in protein crystals

2012 ◽  
Vol 45 (5) ◽  
pp. 1009-1014 ◽  
Author(s):  
Kei Wako ◽  
Kunio Kimura ◽  
Yu Yamamoto ◽  
Takuya Sawaura ◽  
Mengyuan Shen ◽  
...  
Keyword(s):  
Ccd Camera ◽  
Egg White ◽  
Protein Crystals ◽  
Hen Egg White Lysozyme ◽  
X Ray ◽  
Crystal Perfection ◽  
Egg White Lysozyme ◽  
Domain Structures ◽  
Peak Broadening ◽  
Hen Egg White

Digital X-ray topography using an X-ray CCD camera and conventional X-ray topography using X-ray film were used to investigate tetragonal hen egg-white lysozyme (HEWL) crystals. Previously, clear dislocation images of protein crystals were mainly obtained by film methods. Earlier studies of HEWL crystals using an X-ray CCD camera mainly revealed domain structures. In the present study, dislocation images of the same HEWL crystal have been obtained by using conventional X-ray film and a digital X-ray CCD camera. The results demonstrate that digital topography using an X-ray CCD camera is an effective method for characterizing protein crystals. A series of digital topographic images were analyzed by the method developed by Lovelace, Murphy, Pahl, Brister & Borgstahl [J. Appl. Cryst.(2006),39, 425–432]. Sub-peaks and peak broadening originating from dislocations in local rocking curves were observed. Moreover, the crystal perfection was evaluated by mapping the angular positions of the maximums and the full widths at half-maximum of local rocking curves.

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