IVDT_In Vitro Diagnostics Technology

IVD Technology, Spring 2013

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PROCESSING TECHNOLOGIES 80 100 60 Activity % Remaining activity % 100 40 60 40 20 0 80 0 14 28 42 56 70 84 Days at 37ºC 0 6 12 18 24 30 Months at RT Figure 1. Stability of HRP-labeled antibody diluted in an eco-sensible stabilization buffer at room temperature. Remaining activity of Dako Rb a-Hu IgG/HRP diluted 1:2000 in an eco-sensible stabilization buffer (HRP-StabilPLUS) after storage at room temperature (RT) and at 37°C. can be used alone or in combination. Two types of biocides—5-bromo5-nitro-1,3-dioxane (bromonitrodioxane) and isothiazolinone-derivatives— are efective against bacteria, yeast and fungus and are commonly considered to be nontoxic at low concentrations. It should be noted that there is evidence that isothiazolinone-derivatives are rapidly degradable in the environment, but do have toxicity for fsh.1 buffer usage Eco-sensible bufers can be sourced from reagent manufacturers who are committed to environmentally responsible ELISA development by avoiding the use of toxic preservatives without compromising reagent performance or stability (Table I). Tese commercially available eco-sensible bufers can have shelf lives of up to 4 years at 4°C. Since many diagnostic assay kits have a 6- to 12-month shelf life, the use of an eco-sensible bufer will not be a limiting factor for kit production or the determinant of the fnal kit's shelf-life stability. For certain components (i.e., standards and labeled secondary antibody), ecosensible stabilization bufers ofer a practical alternative to lyophilization when the stability of a diagnostic assay at 4°C or room temperature is a critical factor (Figure 1). As an added beneft, eco-sensible bufers enable these components to be provided in a ready-to-use form. name stability Protein-StabilPLUS 2 years at 4°C Stabilization of antibodies or antigen adsorbed to the microwell plate and blocking WellChampion 4 years at 4°C 2. Replace bovine serum albumin Dilution of samples SamplePLUS2 2 years at 4°C HRP-StabilPLUS 4 years at 4°C AP-StabilPLUS 3 years at 4°C Antibody Enhancer 4 years at 4°C In addition to preservatives, many bufers in an ELISA contain blocking agents. A blocking agent's purpose is to disrupt nonspecifc protein binding without cross reacting, exhibiting enzymatic activity, or disrupting specifc protein interactions. Approximately 30 mL of blocking bufer is used per 96-well plate, although blocking agents also can be Stabilization of antibodies, standards, and controls Stabilization of horseradish peroxidase Stabilization of alkaline phosphatase Stabilization and enhancement of horseradish peroxidase Table I. Examples of commercially available eco-sensible buffers for ELISA development and production. These buffers are free from azide, mercury, and other toxic preservatives. 18 IVD TEC HNOLO G Y | SP RIN G 2013 magenta cyan yellow black i v d t e c hnol ogy. com ES236984_IV1305_018.pgs 04.24.2013 23:43 UBM

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