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6.1. INTRODUCTION Quantitative immunoassays constitute an enormous group of assay techniques designed for the selective quantitation of trace levels of low and high molecular weight species in complex biochemical media. In these immunoassay methods, the ligand (antigen or hapten) is almost always the analyte; while the qualitative identi cation of unknown antibodies is often of clinical importance, precise quantitative results are not normally required. In this chapter, the term ligand is used synonymously with hapten and antigen, because secondary binding reactions are not involved. Quantitative immunoassay methods exploit the primary binding of antibody and ligand, that is, the recognition and combination of antibody paratope with antigen epitope, with intrinsic af nity K. Secondary binding interactions do not generally occur, even with macromolecular antigens, because a large excess of either antibody or ligand is usually present. In practice, however, the possibility of secondary interactions should be considered during the development of new immunoassays. The classi cation of immunoassay methods is based on (a) whether they are homogeneous, with no separation step needed prior to measurement, or heterogeneous, where a separation step is required; (b) which species, antibody or antigen, is labeled; and (c) the type of label employed. The ideal label for immunoassay methods has the following properties. It is inexpensive, safe, and simple labeling procedures exist. It is covalently linked to the assay reagent at multiple sites, for high sensitivity. The labeled species is stable. Labeling has a minimal effect on the binding behavior, that is, the labeled and unlabeled reagents behave identically with respect to antibody antigen binding. The label is easily detected using inexpensive instrumentation that is readily automated. Finally, the label should have properties that enable the differentiation of the free and bound forms without requiring a separation step.

Database clients are much easier to recover than NFS clients. When the database server fails, client applications determine that they have lost their server connections and can abort in-progress transactions. Some transactions may have rolled forward when the server recovered, so clients need to reconnect and check on the status of in-flight transactions before restarting them. In

12

Bianalytical Chemistry, by Susan R. Mikkelsen and Eduardo Corton ISBN 0-471-54447-7 Copyright # 2004 John Wiley & Sons, Inc.

general, database client libraries handle reconnection, transaction restart, and recovery gracefully When using an HA pair, the IP address and hostname used for the data service migrate to the secondary server, so clients reconnect to the right machine The generic case is the easy case It s possible for stored procedures or triggers to fail on an otherwise normally functioning database These failures may be due to internal database problems; errors in the logic, external problems such as a full log device or lack of data table space, or a lack of available memory on the server Client applications must check the return values of stored procedures, looking for all possible failure modes and exception conditions Also watch out for longrunning transactions.

The examples that follow are all taken from genuine, observed facts. For reasons of confidentiality, the business lines and some elements have been changed. 11.1 Rationalizing bond purchase order management (banking) 11.1.1. The context Bermano Bank, a subsidiary of a large European bank, negotiates financial and derivative instrument products in different markets, ranging from highly regulated domestic markets, and extending to unquoted and international markets. This diversity of instruments and markets is reflected in the diversity of the IT applications required in both the front-office and back-office to manage a variety of bonds. For historical reasons, as in all information systems, integration between these applications was carried out one step at a time, by creating point-to-point links, with all the consequences already mentioned in this work: frequent manual intervention to process errors, high maintenance costs throughout the system, technical touchups to make up for missing functions and correct breakdowns in processing purchase orders. Moreover, integration between certain applications relies on batches, which add complementary delays to the process of purchasing bonds. The existing system is summarized by Figure 11.1.

obtained at each protein concentration indicate that the protein does not selfassociate The sedimentation coef cient (s0 ) was calculated as 708 009 S, 20;w from which the frictional ratio of SAT was found to be 153; this value is high when compared with the expected value for a globular protein ($ 115), supporting the hypothesis that SAT subunits are not organized in a compact motif This hypothesis was con rmed using other techniques The authors conclude that the fundamental building block is probably a trimer, with SAT arranged as a pair of loosely staked trimers 1392 Isolation of Retroviruses by Self-Generated Gradients18 Retroviruses are viruses in which the genetic material is RNA Several retroviruses are related with human diseases, like in uenza or HIV.

Clients that set timeouts (or deadman timers) for some transactions may inadvertently abort transactions that are blocked by a large SELECT or other transaction that consumes most of the database engine s cycles Application developers may be tempted to break up the single large transaction into several smaller ones, performing part of the work in each transaction This attempt to break the logjam works well if you are certain that the other transactions are lookup- or read-oriented, and won t change data that impacts parts of the broken-up transaction that occur later in the overall timeline Part of the advantage of a single, large transaction is that you re guaranteed to have the database state remain consistent during the course of the entire work unit You need to manage your recovery time exposure against the smallest logical unit of work done by the application.