Construction and characterization of a novel recombinant single-chain variable fragment antibody against White Spot Syndrome Virus from shrimp

An antibody phage display library against White Spot Syndrome Virus (WSSV) was constructed. After four rounds of panning against WSSV, 192 out of 480 clones displayed WSSV binding activity. One of the positive clones, designated A1, had relatively higher activity specifically binding to WSSV A1-soluble, single-chain fragment variable (scFv) antibody has an affinity constant (K-aff) of 2.02 +/- 0.42 x 10(9) M-1. Dot blot assays showed that A1-soluble scFv could detect WSSV directly from shrimp hemolymph after 24-h feeding infection by WSSV. A1 scFv has potential for the development of a cheap, simple and sensitive diagnostic kit for WSSV in the field. (C) 2003 Elsevier Science B.V. All rights reserved.

Detection of Myxobolus rotundus (Myxozoa : Myxosporea) in skin mucus of crucian carp Carassius auratus auratus using a monoclonal antibody

Diagnosis of myxosporean Myxobolus rotundus infection was conducted by examining skin mucus from the infected crucian carp Carassius auratus auratus with a monoclonal antibody, MAb 2D12, raised previously against the parasite. A positive reaction was observed in skin mucus collected from infected fish, and spores and pre-spore stages of the parasite were identified by the MAb 2D12. It was also demonstrated that M. rotundus infection can be successfully detected by a simple method, enzyme-linked immunosorbent assay (ELISA), and that skin mucus collected from infected fish skin had a significantly higher optical density (OD) value than that from uninfected fish.

Isolation by phage display and characterization of a single-chain antibody specific for O-6-methyldeoxyguanosine

New approaches of making single chain Fv antibodies against O-6-methyl-2'-deoxyguanosine (O(6)MdG) have been demonstrated by using the phage antibody display system. Using O(6)MdG as an antigen, 21 positive clones were identified by ELISA from this library, one of which, designated H3, specifically binds to O(6)MdG with high affinity. The H3 scFv antibody has an affinity constant (K-aff) of 5.94 x 10(11)(mol/L)(-1). H3 scFv has been successfully used to detect O-6 MdG in DNA hydrolyses from yeast or E. coli cells treated with a DNA methylating agent. To our knowledge, this is the first report of the selection of a specific scFv against DNA adducts. The results demonstrate the potential applications of the phage display technology for the detection of DNA lesions caused by mutagens and carcinogens.

Antibody response of carp, Cyprinus carpio to the cestode, Bothriocephalus acheilognathi

The humoral antibody response and the number of pronephric antibody-secreting cells were examined in naturally Bothriocephalus acheilognathi-infected carp. Cyprinus carpio, and in those injected intraperitoneally with an extract of the cestode. In the extract-injected fish, specific antibody was detected 3 weeks after a second injection given 2 weeks after the primary injection, and antibody levels persisted for more than 200 days. A third injection also enhanced the antibody level in the extract-injected carp. The numbers of antibody-secreting cells were significantly higher in carp injected 3 times with the extract than in the control. In naturally-infected fish, the serum antibody levels and the number of pronephric antibody-secreting cells were higher in infected fish than in uninfected individuals although this difference was not statistically significant. The relevance of these results to immune protection against infection is discussed.

Hole concentration test of p-type GaN by analyzing the spectral response of p-n(+) structure GaN ultraviolet photodetector

A new method to test the hole concentration of p-type GaN is proposed, which is carried out by analyzing the spectral response of p-n(+) structure GaN ultraviolet photodetector. It is shown that the spectral response of the photodetector changes considerably with reversed bias. It is found that the difference between photodetector's quantum efficiency at two wavelengths, i.e. 250 and 361 nm, varies remarkably with increasing reversed bias. According to the simulation calculation, the most characteristic change occurs at a reversed voltage under which the p-GaN layer starts to be completely depleted. Based on this effect the carrier concentration of p-GaN can be derived.

Fracture properties of silicon carbide thin films by bulge test of long rectangular membrane

This paper reports the mechanical properties and fracture behavior of silicon carbide (3C-SiC) thin films grown on silicon substrates. Using bulge testing combined with a refined load-deflection model of long rectangular membranes, which takes into account the bending stiffness and prestress of the membrane material, the Young's modulus, prestress, and fracture strength for the 3C-SiC thin films with thicknesses of 0.40 and 1.42 mu m were extracted. The stress distribution in the membranes under a load was calculated analytically. The prestresses for the two films were 322 +/- 47 and 201 +/- 34 MPa, respectively. The thinner 3C-SiC film with a strong (111) orientation has a plane-gstrain moduli of 415 +/- 61 GPa, whereas the thicker film with a mixture of both (111) and (110) orientations exhibited a plane-strain moduli of 329 +/- 49 GPa. The corresponding fracture strengths for the two kinds of SiC films were 6.49 +/- 0.88 and 3.16 +/- 0.38 GPa, respectively. The reference stresses were computed by integrating the local stress of the membrane at the fracture over edge, surface, and volume of the specimens and were fitted with Weibull distribution function. For the 0.40-mu m-thick membranes, the surface integration has a better agreement between the data and the model, implying that the surface flaws are the dominant fracture origin. For the 1.42-mu m-thick membranes, the surface integration presented only a slightly better fitting quality than the other two, and therefore, it is difficult to rule out unambiguously the effects of the volume and edge flaws.

Choice of calibration equations in calibrating microwave test fixtures

The error theory of linear equation system has been applied to the calibration procedure of microwave network analyser in this article. A new explanation for the choice of the linear calibration equations is proposed and a general principle for choosing calibration equations is presented. The method can also be used to predict the occurrence of the problem of frequency limitation at some periodic frequencies. This principle is employed to the thru-short-delay (TSD) method and the solution using the chosen equations gives the most accurate results. A good agreement between the theory and the experiment has been obtained.

An improved TM method for full two-port calibration of the asymmetric test fixtures

Three known standards, including at least one transmission standard, are normally required for the full two-port calibration of test fixtures. Based on the triple-through method, a new general-purpose calibration procedure using only one known reflection standard is proposed in this paper. The experimental results show that our method call provide a simple and accurate approach to fall two-port calibration of the asymmetric test fixtures. (c) 2005 Wiley Periodicals, Inc.

Frequency limitation in the calibration of microwave test fixtures

The problem of frequency limitation arising from the calibration of asymmetric and symmetric test fixtures has been investigated. For asymmetric test fixtures, a new algorithm based on the thru-short-match (TSM) method is outlined. It is found that the conventional TSM method does not have any inherent frequency limitation, but using the same procedure with an unknown match may lead to the said problem. This limitation can be avoided by using a different algorithm. The various calibration methods for symmetric test fixtures using known standards are also discussed and the origin of the frequency limitation is identified. Several ways in avoiding the problem are proposed. There is good agreement between the theories and experimental data.

Two-port calibration of test fixtures with different test ports

The open-short-load (OSL) method is very simple and widely used for one-port test fixture calibration. In this paper, this method is extended, for the first time, to the two-port calibration of test fixtures with different test ports. The problem of phase uncertainty arising in this application has been solved. The comparison between our results and those obtained with the SOLT method shows that the method established is accurate for practical applications. (C) 2002 Wiley Periodicals, Inc.

New algorithms of the TSM and TOM methods for calibrating microwave test fixtures

Based on the conventional through-short-match (TSM) method, an improved TSM method has been proposed in this Letter. This method gives an analytical solution and has almost all the advantages of conventional TSM methods. For example, it has no phase uncertainty and no bandwidth limitation. The experimental results show that the accuracy can be significantly improved with this method. The proposed theory can be applied to the through-open-match (TOM) method. (C) 2002 Wiley Periodicals. Inc.

Phase uncertainty in calibrating microwave test fixtures

The problem of phase uncertainty arising in calibration of the test fixtures is investigated in this paper, It is shown that the problem exists no matter what kinds of calibration standards are used. It is also found that there is no need to determine the individual S-parameters of the test fixtures. In order to eliminate the problem of phase uncertainty, three different precise (known) reflection standards or one known reflection standard plus one known transmission standard should be used to calibrate symmetrical test fixtures. For the asymmetrical cases, three known standards, including at least one transmission standard, should be used. The thru-open-match (TOM) and thru-short-match (TSM) techniques are the simplest methods, and they have no bandwidth limitation. When the standards are imprecise (unknown), it is recommended to use any suitable technique, such as the thru-reflect-line, line-reflect-line, thru-short-delay, thru-open-delay,line-reflect-match, line-reflect-reflect-match, or multiline methods, to accurately determine the values of the required calibration terms and, in addition, to use the TOM or TSM method with the same imprecise standards to resolve the phase uncertainty.

Correlation of calibration equations for test fixtures

This paper begins from the thru-short-open (TSO) and thru-line-match (TLM) methods to investigate the correlation of the calibration equations of these two methods, The relations among the measurements with the corresponding standards are obtained. It is found that the line standard with zero length can be used instead of ideal open and short, in case that two test fixtures are symmetrical. For asymmetrical fixtures, the measurements with the standards line, open and short are related at certain frequencies, and the matched load can be replaced by the line standards. The relations established are used to test short and match standards and analyze the freqPuency limits of the TSO method, Good agreement between theory and experiment is obtained, It is found that the TSO method becomes very poor when the insertion phase of the thru standard is near n pi/4, and this method has a lower frequency limit. The TLM method is found unsuitable for calibrating asymmetrical fixtures.

Fracture properties of PECVD silicon nitride thin films by long rectangular memrane bulge test

Bulge test combined with a refined load-deflection model for long rectangular membrane was applied to determine the mechanical and fracture properties of PECVD silicon nitride (SiNx) thin films. Plane-strain modulus E-ps prestress s(0), and fracture strength s(max) of SiNx thin films deposited both on bare Si substrate and on SiO2-topped Si substrate were extracted. The SiNx thin films on different substrates possess similar values of E-ps and s(0) but quite different values of s(max). The statistical analysis of fracture strengths were performed by Weibull distribution function and the fracture origins were further predicted.