Selection of Diethylstilbestrol-Specific Single-Chain Antibodies from a Non-Immunized Mouse Ribosome Display Library

Single chain variable fragments (scFvs) against diethylstilbestrol (DES) were selected from the splenocytes of non-immunized mice by ribosome display technology. A naive library was constructed and engineered to allow in vitro transcription and translation using an E. coli lysate system. Alternating selection in solution and immobilization in microtiter wells was used to pan mRNA-ribosome-antibody (ARM) complexes. After seven rounds of ribosome display, the expression vector pTIG-TRX containing the selected specific scFv DNAs were transformed into Escherichia coli BL21 (DE3) for expression. Twenty-six positive clones were screened and five clones had high antibody affinity and specificity to DES as evidenced by indirect competitive ELISA. Sequence analysis showed that these five DES-specific scFvs had different amino acid sequences, but the CDRs were highly similar. Surface plasmon resonance (SPR) analysis was used to determine binding kinetics of one clone (30-1). The measured KD was 3.79 µM. These results indicate that ribosome display technology can be used to efficiently isolate hapten-specific antibody (Ab) fragments from a naive library; this study provides a methodological framework for the development of novel immunoassays for multiple environmental pollutants with low molecular weight detection using recombinant antibodies

Problema de programação da produção um esquema de classificação

Muitas vezes, não é simples encontrar uma classificação exata para os problemas de programação, não somente porque existem diferentes versões para um dado problema, mas, porque vários procedimentos para uma questão particular, são caracterizados por premissas diferentes e limitações de aplicação dos modelos desenvolvidos. O objetivo deste artigo é delinear uma classificação ampla que permita estabelecer o sentido, direção e perspectiva de pesquisas conduzidas na área. O trabalho não tem a intenção de dar um levantamento exaustivo da literatura de programação da produção, que pode ser encontrado em vários outros trabalhos de revisão.<br>It is the purpose of this article to review the various solutions that have been proposed for the production scheduling problem. An attempt is made to give a classification scheme to categorize the existing procedures that allow to point out potential future courses of development. Emphasis is placed on the basic assumptions involved in each production sequencing problem rather than to approaches used to obtain a solution