Surfactant‐Induced Amorphous Aggregation of Tobacco Mosaic Virus Coat Protein: A Physical Methods Approach

Panyukov, Yuliy V.; Nemykh, Maria A.; Добров, Е. Н.; Drachev, V. A.

doi:10.1002/mabi.200700145

Cited by 13 publications

(9 citation statements)

References 57 publications

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“…Particles are first denatured in 0.1% SDS by heating at 95°C for 10 minutes, and samples are then treated with 1% NP-40 in order to sequester the SDS (Panyukov and others 2008) and prevent it from interfering with subsequent steps. Preparations can then be directly digested with restriction endonuclease prior to quantitation with qPCR using primer set 2 (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Systemic Errors in Quantitative Polymerase Chain Reaction Titration of Self-Complementary Adeno-Associated Viral Vectors and Improved Alternative Methods

Fagone

Wright

Nathwani

et al. 2012

Human Gene Therapy Methods

100

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Self-complementary AAV (scAAV) vector genomes contain a covalently closed hairpin derived from a mutated inverted terminal repeat which connects the two monomer single stranded genomes into a head-to-head or tail-to-tail dimer. We found that during quantitative PCR (qPCR) this structure inhibits the amplification of proximal amplicons and causes the systemic underreporting of copy number by as much as 10-fold. We show that cleavage of scAAV vector genomes with restriction endonuclease to liberate amplicons from the covalently closed terminal hairpin restores quantitative amplification, and we implement this procedure in a simple, modified qPCR titration method for scAAV vectors. Additionally, we developed and present an AAV genome titration procedure based on gel electrophoresis that requires minimal sample processing and has very low inter assay variability, and as such is well suited for the rigorous quality control demands of clinical vector production facilities.

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Section: Resultsmentioning

confidence: 99%

Systemic Errors in Quantitative Polymerase Chain Reaction Titration of Self-Complementary Adeno-Associated Viral Vectors and Improved Alternative Methods

Fagone

Wright

Nathwani

et al. 2012

Human Gene Therapy Methods

100

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show abstract

“…Moreover, SDS has been used to prevent the transmission of HIV during sexual intercourse (25). In addition, Song and others (2010) reported that SDS, NP-40, and Triton X-100 were able to reduce the infectivity of hepatitis C virus (46), whereas it has been reported that Triton X-100 was able to partially denature the coat protein of tobacco mosaic virus (TMV) and then induce aggregation of this coat protein (38). However, the effectiveness of surfactants inactivating food-borne viruses is less understood.…”

Section: Discussionmentioning

confidence: 99%

Enhanced Removal of a Human Norovirus Surrogate from Fresh Vegetables and Fruits by a Combination of Surfactants and Sanitizers

Predmore

Lǐ

2011

Appl Environ Microbiol

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Fruits and vegetables are major vehicles for transmission of food-borne enteric viruses since they are easily contaminated at pre-and postharvest stages and they undergo little or no processing. However, commonly used sanitizers are relatively ineffective for removing human norovirus surrogates from fresh produce. In this study, we systematically evaluated the effectiveness of surfactants on removal of a human norovirus surrogate, murine norovirus 1 (MNV-1), from fresh produce. We showed that a panel of surfactants, including sodium dodecyl sulfate (SDS), Nonidet P-40 (NP-40), Triton X-100, and polysorbates, significantly enhanced the removal of viruses from fresh fruits and vegetables. While tap water alone and chlorine solution (200 ppm) gave only <1.2-log reductions in virus titer in all fresh produce, a solution containing 50 ppm of surfactant was able to achieve a 3-log reduction in virus titer in strawberries and an approximately 2-log reduction in virus titer in lettuce, cabbage, and raspberries. Moreover, a reduction of approximately 3 logs was observed in all the tested fresh produce after sanitization with a solution containing a combination of 50 ppm of each surfactant and 200 ppm of chlorine. Taken together, our results demonstrate that the combination of a surfactant with a commonly used sanitizer enhanced the efficiency in removing viruses from fresh produce by approximately 100 times. Since SDS is an FDA-approved food additive and polysorbates are recognized by the FDA as GRAS (generally recognized as safe) products, implementation of this novel sanitization strategy would be a feasible approach for efficient reduction of the virus load in fresh produce.

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“…It was shown that Triton X -100 can induce a partial denaturation of the tobacco mosaic virus coat protein, and surfactant -protein complexes aggregate though hydrophobic interactions. 233 …”

Section: Nonionic Surfactantsmentioning

confidence: 99%

External Factors Affecting Protein Aggregation

Wang

Speaker

2010

Aggregation of Therapeutic Proteins

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The behavior of biotherapeutic proteins is signifi cantly different from small molecule drugs both in liquid and solid states, especially in terms of physical stability. One such property is the high tendency of protein molecules to aggregate under a variety of conditions. The aggregation tendency is arguably the most common and troubling manifestation of protein instability during the development of protein biotherapeutics. 1 Protein aggregates usually exhibit either reduced or, in many cases, no biological activity. 2 -5 A clear correlation was established between loss of recombinant factor VIII (rFVIII ) and its degree of aggregation as shown in Fig. 4.1 . 6 To achieve effective prevention or inhibition of protein aggregation, it is of paramount importance to understand all the possible factors that affect or control the protein aggregation process.Many factors have been identifi ed and reported in the literature affecting the process and rate of protein aggregation. These factors can be roughly divided into two major categories: internal (protein structure related) and external (protein environment related). Chapter 3 in this book has focused extensively on factors that belong to the fi rst category. This chapter focuses on the external factors that affect the process and rate of protein aggregation. To facilitate discussion, these external factors are further divided into the following sections: (1) temperature; (2) solution conditions and composition (pH, buffer type and concentration, ionic strength, excipients and level, protein concentration, metal ions, denaturing and reducing agents, impurities, organic solvents, containers/closures, sources of proteins, sample treatment, and analytical methodologies); (3) processing steps (fermentation/expression,

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Surfactant‐Induced Amorphous Aggregation of Tobacco Mosaic Virus Coat Protein: A Physical Methods Approach

Cited by 13 publications

References 57 publications

Systemic Errors in Quantitative Polymerase Chain Reaction Titration of Self-Complementary Adeno-Associated Viral Vectors and Improved Alternative Methods

Systemic Errors in Quantitative Polymerase Chain Reaction Titration of Self-Complementary Adeno-Associated Viral Vectors and Improved Alternative Methods

Enhanced Removal of a Human Norovirus Surrogate from Fresh Vegetables and Fruits by a Combination of Surfactants and Sanitizers

External Factors Affecting Protein Aggregation

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