Question

Which of the following precautions must not be followed while performing assays?

(a) The substrates, buffers etc., should be of high purity

(b) Enzyme preparation should as pure as possible

(c) The probe must be tiny and biocompatible

(d) Stability of the enzyme during the time taken by assay

I got this question in my homework.

The query is from Continuous, Discontinuous and Coupled Assay in portion Enzyme Assay & Enzyme Catalysis of Enzyme Technology

Answer 1

Right answer is (c) The probe must be tiny and biocompatible

Best explanation: “The probe must be tiny and biocompatible.” This is feature of biosensor, and not precautions to be followed. Following are the precautions that must not be followed while performing assays:

• The substrates, buffers etc., should be of high purity as the contaminants may affect the activity of the enzyme. For example: Certain NAD^+ preparations contain inhibitor of dehydrogenases.

• Enzyme preparation should as pure as possible so that any other compounds or enzyme will not interfere with assay

• Stability of the enzyme during the time taken by assay to complete the reaction.

• pH, temperature etc., affects the activity of the enzyme, and hence has to be maintained by use of buffers, thermostable bath etc.

• It should be ensured that once steady state is reached, the rate of reaction is constant over a period of interest and proportional to the amount of enzyme added.