Recent developments in non-enzymatic (bio)sensors for detection of pesticide residues: Focusing on antibody, aptamer and molecularly imprinted polymer.

The utilization of pesticides has been increased in recent years due to population growth and increasing urbanization. The constant use of pesticides has resulted in contamination of the environment and agricultural products with serious human health concerns associated with their use. Therefore, detection and quantification of pesticides by sensitive and selective methods is highly required in food safety management. Traditional detection methods cannot realize highly sensitive, selective and on-site detection, which limits their application. (Bio)sensors and (bio)assays are emerging tools with unique properties such as rapid, sensitive, efficient and portable detection. Among them, enzyme-based biosensors have been widely developed and some have even been commercialized. However, they suffer from some limitations such as instability and low reproducibility that originate from the nature of enzyme. Non-enzymatic (bio)sensors overcome the current limitations of enzyme-based detection methods and provide great potential for efficient, highly sensitive and low-cost detection assays using smart and miniaturized devices. In this study, we provide an overview of recent advances and new trends in optical and electrochemical non-enzymatic (bio)sensors for the detection of pesticides by focusing on antibody, aptamer and molecularly imprinted polymer (MIP) as recognition elements. Performance, advantages and drawbacks of the developed (bio)sensors are discussed well. The main advantage these recognition elements is their stability over an extended period of time compared to the enzymes. Furthermore, the combination of nanomaterials in these (bio)sensors can significantly improve their performance.

Experimental investigation of the frequency and substitution dependence of negative phi-values in two-state proteins.

Negative phi-values, which arise, for example, when a mutation stabilizes the folding transition state while destabilizing the native state, have been the focus of significant theoretical interest. Here we survey the experimental folding kinetics literature to ascertain the frequency with which negative phi-values occur in two-state proteins and describe the detailed experimental characterization of a negative phi-value previously reported to be among the most statistically significant. We find that, while almost 9% of more than 500 reported phi-values (from a set of 16, well-characterized two-state proteins) fall below zero, many of these do not represent statistically significant observations. For example, only 6% of the phi-values for which estimates of precision are available fall even one reported "error bar" below zero, and only 4% are simultaneously negative, significant at this level and associated with free energy changes at or above 2.5 kJ/mol (below which phi-value analysis is widely considered unreliable). Moreover, given the asymmetric distribution of phi-values around zero and given that reported error bars may significantly underestimate true confidence intervals, the actual number of negative phi-values may be much smaller still. We have also performed detailed characterization of one of the most statistically significant negative phi-values reported in the literature to date, the V55F mutant of FynSH3. We find that substitution of the wild-type valine to other hydrophobic residues often increases folding rates without significantly altering folding free energy. This in turn leads to poorly defined phi-values, some of which are formally negative but only one or two of which fall statistically significantly below zero. In contrast, substitution to polar residues significantly destabilizes both the transition and native states, generally producing small but statistically significant positive phi-values of approximately 0.1. Thus, unlike other previously characterized phi-values, the negative phi-value associated with position 55 of the FynSH3 domain appears to be strongly dependent on the substitution employed to measure it, suggesting that subtlety will be required in order to develop a theoretical model of such behavior.