The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis.

A member of a specific binding pair (sbp) is identified by expressing DNA encoding a genetically diverse population of such sbp members in recombinant host cells in which the sbp members are displayed in functional form at the surface of a secreted recombinant genetic display package (rgdp) containing DNA encoding the sbp member or a polypeptide component thereof, by virtue of the sbp member or a polypeptide component thereof being expressed as a fusion with a capsid component of the rgdp. The displayed sbps may be selected by affinity with a complementary sbp member, and the DNA recovered from selected rgdps for expression of the selected sbp members. Antibody sbp members may be thus obtained, with the different chains thereof expressed, one fused to the capsid component and the other in free form for association with the fusion partner polypeptide. A phagemid may be used as an expression vector, with said capsid fusion helping to package the phagemid DNA. Using this method libraries of DNA encoding respective chains of such multimeric sbp members may be combined, thereby obtaining a much greater genetic diversity in the sbp members than could easily be obtained by conventional methods.

Methods for producing members of specific binding pairs

http://theory.bio.uu.nl/immbio/Marks91.pdf

By-passing immunization: Human antibodies from V-gene libraries displayed on phage

Molecular cloning and expression of the fas ligand, a novel member of the tumor necrosis factor family

Tens of millions of short peptides can be easily surveyed for tight binding to an antibody, receptor or other binding protein using an "epitope library." The library is a vast mixture of filamentous phage clones, each displaying one peptide sequence on the virion surface. The survey is accomplished by using the binding protein to affinity-purify phage that display tight-binding peptides and propagating the purified phage in Escherichia coli. The amino acid sequences of the peptides displayed on the phage are then determined by sequencing the corresponding coding region in the viral DNA's. Potential applications of the epitope library include investigation of the specificity of antibodies and discovery of mimetic drug candidates.

https://science.sciencemag.org/content/sci/249/4967/386.full.pdf

Searching for Peptide Ligands With an Epitope Library

E. coli can be transformed to extremely high efficiencies by subjecting a mixture of cells and DNA to brief but intense electrical fields of exponential decay waveform (electroporation). We have obtained 10(9) to 10(10) transformants/micrograms with strains LE392 and DH5 alpha, and plasmids pUC18 and pBR329. The process is highly dependent on two characteristics of the electrical pulse: the electric field strength and the pulse length (RC time constant). The frequency of transformation is a linear function of the DNA concentration over at least six orders of magnitude; and the efficiency of transformation is a function of the cell concentration. Most of the surviving cells are competent with up to 80% transformed at high DNA concentration. The mechanism does not appear to include binding of the DNA to the cells prior to entry. Possible mechanisms are discussed and a simple procedure for the practical use of this technique is presented.

High efficiency transformation of E.coli by high voltage electroporation

Individual temperature control in multiple reactions performed simultaneously in a spatial array such as a multi-well plate is achieved by thermoelectric modules with individual control, with each module supplying heat to or drawing heat from a single region within the array, the region containing either a single reaction vessel or a group of reaction vessels.

Localized temperature control for spatial arrays of reaction media

An arc detector for detecting electrical arcs across a break in a line carrying a current in an electrical system. Normally, the line carries current, but when the line is broken, current doesn't flow until an arc occurs across the break. The arc detector detects the break using a current sensor for measuring the current, a filter for filtering out unwanted frequencies, an amplifier for amplifying the remaining frequencies into a substantially square wave, a counter for counting square wave transitions, and a latch to hold a detection signal at an output node when the counter counts a preset number of square wave transitions without being reset by a reset switch or a periodic oscillator.

Arc detection system

A method is provided for measuring a response to a stimulus of a plurality of samples spots of a sample using a measuring system having a measurement range to generate an image of the sample in digital space. The method includes, for each sample, while measuring the response, varying the stimulus to include at least one stimulus value where the measured response corresponds to a value in an intermediate portion of the measuring range, and storing a value of the measured response that corresponds to a value in the intermediate portion of the measurement range, and the stimulus value that produced that value of the measured response.

Imaging method and apparatus

The present invention provides apparatus and method for simply and efficiently improving prior art transfection systems. The present invention is a transfection high-voltage controller that provides features for measuring actual load resistance, and compensating for any effects of a limit impedance in series with the load. Additionally, the transfection controller of the present invention measures actual effective capacitance of a storage capacitor, or capacitors, and through use of an expansion module, effectively provides the user with a set of user-selectable capacitors having an effective ±10% tolerance when the tolerance of the individual electrolytic capacitors is ±20%. The preferred embodiment incorporates two sections. One section of the system handles 200-2500V and another section handles low voltages (50-500V). The transfection controller includes an improved hi-joule switch that operates in a low-voltage, high-current mode with a low dropout voltage (for use with the 500V (maximum) electrolytic capacitors. Further, additional protective circuitry in the form of a crowbar circuit, will not only prevent charging of the storage capacitor unless a safety switch is operated, charge from the capacitor is shunted to ground should the charging operation be interrupted, or if power is removed from the transfection system.

Charles W. Ragsdale

Papers

High efficiency transformation of E.coli by high voltage electroporation

Localized temperature control for spatial arrays of reaction media

Arc detection system

Imaging method and apparatus

Transfection high-voltage controller