Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity

One of existing strategies to engineer active antibody is to link VH and VL domains via a linker peptide. How the composition, length, and conformation of the linker affect antibody activity, however, remains poorly understood. In this study, a dual approach that coordinates molecule modeling, biological measurements, and affinity evaluation was developed to quantify the binding activity of a novel stable miniaturized anti-CD20 antibody or singlechain fragment variable (scFv) with a linker peptide. Upon computer-guided homology modeling, distance geometry analysis, and molecular superimposition and optimization, three new linker peptides PT1, PT2, and PT3 with respective 7, 10, and 15 residues were proposed and three engineered antibodies were then constructed by linking the cloned VH and VL domains and fusing to a derivative of human IgG1. The binding stability and activity of scFv-Fc chimera to CD20 antigen was quantified using a micropipette adhesion frequency assay and a Scatchard analysis. Our data indicated that the binding affinity was similar for the chimera with PT2 or PT3 and ~24-fold higher than that for the chimera with PT1, supporting theoretical predictions in molecular modeling. These results further the understanding in the impact of linker peptide on antibody structure and activity.

Absolute measurement of roughness and lateral-correlation length of random surfaces by use of the simplified model of image-speckle contrast

We present a method of image-speckle contrast for the nonprecalibration measurement of the root-mean-square roughness and the lateral-correlation length of random surfaces with Gaussian correlation. We use the simplified model of the speckle fields produced by the weak scattering object in the theoretical analysis. The explicit mathematical relation shows that the saturation value of the image-speckle contrast at a large aperture radius determines the roughness, while the variation of the contrast with the aperture radius determines the lateral-correlation length. In the experimental performance, we specially fabricate the random surface samples with Gaussian correlation. The square of the image-speckle contrast is measured versus the radius of the aperture in the 4f system, and the roughness and the lateral-correlation length are extracted by fitting the theoretical result to the experimental data. Comparison of the measurement with that by an atomic force microscope shows our method has a satisfying accuracy. (C) 2002 Optical Society of America.

Influence of Coulomb potential for photoionization of H atoms in an elliptically polarized laser field: Velocity gauge versus length gauge

We theoretically study the influence of Coulomb potential for photoionization of hydrogen atoms in an intense laser field with elliptical polarization. The total ionization rates, photoelectron energy spectra, and photoelectron angular distributions are calculated with the Coulomb-Volkov wave functions in the velocity gauge and compared with those calculated in the length gauge as well as those calculated with the Volkov wave functions. By comparing the results obtained by the Coulomb-Volkov and Volkov wave functions, we find that for linear polarization the influence of Coulomb potential is obvious for low-energy photoelectrons, and as the photoelectron energy and/or the laser intensity increase, its influence becomes smaller. This trend, however, is not so clear for the case of elliptical polarization. We also find that the twofold symmetry in the photoelectron angular distributions for elliptical polarization is caused by the cooperation of Coulomb potential and interference of multiple transition channels. About the gauge issue, we show that the difference in the photoelectron angular distributions obtained by the velocity and length gauges becomes rather obvious for elliptical polarization, while the difference is generally smaller for linear polarization.

Spatial longitudinal coherence length of a thermal source and its influence on lensless ghost imaging

The spatial longitudinal coherence length (SLCL), which is determined by the size of and the distance from the source, is introduced to investigate the longitudinal resolution of lensless ghost imaging. Its influence is discussed quantitatively by simulation. The discrepancy of position sensitivity between Scareelli et al. [Appl. Phys. Lett. 88, 061106 (2006)] and Basano and Ottonello [Appl. Phys. Lett. 88, 091109 (2006)] is clarified. (C) 2008 Optical Society of America.

The photorefractive sensitivity and the effective electron transport length in two-centre holographic recording

In this paper the photorefractive sensitivity defined for single-centre holographic recording is modified to adapt two-centre holographic recording. Based on the time analytic solution of Kukhtarev equations for doubly doped crystals, the analytical expression of photorefractive sensitivity is given. For comparison with single-centre holographic recording and summing the electron competition effects between the deeper and shallower traps, an effective electron transport length is proposed, which varies with the intensity ratios of recording light to sensitive light. According to analyses in this paper, the lower photorefractive sensitivity in two-centre holographic recording is mainly due to the lower concentration of unionized dopants in the shallower centre and the lower effective electron transport length.

Design of a highly stable, high-conversion-efficiency pumping source for optical parametric amplifier by extending efficient crystal length

The objective of this study is to improve the stability of pumping source of optical parametric amplifier. Analysis by simulation leads to the conclusion that the stability of the second harmonic can be improved by using properly the intensity of fundamental light and corresponding length of the crystal. By the method of the noncollinear two-pass second harmonic or the tandem second harmonic, the efficient crystal length is extended to a proper value, and the stability of the second harmonic output has been improved two times more than that for the fundamental light, and the conversion-efficiency is about 70% in experiment. When the variation of the fundamental light is about 10%, the variation of the second harmonic intensity has been controlled within 5%. (c) 2006 Elsevier Ltd. All rights reserved.

Passive Q-switching of short-length Tm3+-doped silica fiber lasers by polycrystalline Cr2+:ZnSe microchips

We demonstrate passive Q-switching of short-length double-clad Tm3+-doped silica fiber lasers near 2 mu m pumped by a laser diode array (LDA) at 790 nm. Polycrystalline Cr2+:ZnSe microchips with thickness from 0.3 to 1 mm are adopted as the Q-switching elements. Pulse duration of 120 ns, pulse energy over 14 mu] and repetition rate of 53 kHz are obtained from a 5-cm long fiber laser. As high as 530 kHz repetition rate is achieved from a 50-cm long fiber laser at similar to 10-W pump power. The performance of the Q-switched fiber lasers as a function of fiber length is also analyzed. (c) 2008 Elsevier B.V. All rights reserved.

The Homology and Relationship of Human (Homo sapiens) Chromosomes 1, 19 and Dusky Langur (Trachypithacus obscurus) Chromosomes 6, 8 Demonstrated with Chromosome Painting

http://www.jstage.jst.go.jp/

A COMPARATIVE-STUDY OF KARYOTYPES OF MUNTJACS BY CHROMOSOME PAINTING

We have used a combination of chromosome sorting, degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), chromosome painting and digital image capturing and processing techniques for comparative chromosome analysis of members of the genus Muntiacus. Chromosome-specific ''paints'' from a female Indian muntjac were hybridised to the metaphase chromosomes of the Gongshan, Black, and Chinese muntjac by both single and three colour chromosome painting. Karyotypes and idiograms for the Indian, Gongshan, Black and Chinese muntjac were constructed, based on enhanced 4', 6-diamidino-2-phenylindole (DAPI) banding patterns. The hybridisation signal for each paint was assigned to specific bands or chromosomes for all of the above muntjac species. The interspecific chromosomal homology was demonstrated by the use of both enhanced DAPI banding and comparative chromosome painting. These results provide direct molecular cytogenetic evidence for the tandem fusion theory of the chromosome evolution of muntjac species.

Evolution of the black muntjac (Muntiacus crinifrons) karyotype revealed by comparative chromosome painting

The black muntjac (Muntiacus crinifrons) has an unusual karyotype of 2n = 8 in females and 2n = 9 in males. We have studied the evolution of this karyotype by hybridising chromosome-specific paints derived from flow-sorted chromosomes of the Chinese muntjac (M. reevesi, 2n = 46) to chromosomes of the black muntjac. The hybridisation pattern allowed us to infer chromosomal homologies between these two species. Tandem and centromeric fusions, reciprocal translocations, and insertions are involved in the reduction of the diploid number from 2n = 46 to 2n = 8, 9. The painting patterns further show complex chromosomal rearrangements in the male black muntjac which involve more than half the karyotype, including both sex chromosomes. Since early meiosis is reported to be normal without any visible inversion loops of the synaptonemal complex, the observed chromosomal rearrangements would lead to heterosynapsis and, therefore, leave a large fraction of the male black muntjac karyotype balanced between the two sexes.