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Papers by Marina Feroggio

Research paper thumbnail of LC-MS Method for the Quantitation of Two Monoclonal Antibodies by Multiple Signature Peptides in Monkey Serum

Journal of Analytical & Bioanalytical Techniques, 2015

Evaluation of the in-vivo concentration of monoclonal antibody (mAb) mixtures is a challenging ta... more Evaluation of the in-vivo concentration of monoclonal antibody (mAb) mixtures is a challenging task. Here we report the application of an LC-MS bioanalytical method to quantify in monkey serum the Sym004, an equimolar mixture of two monoclonal antibodies, 992 mAb and 1024 mAb. This method has been assessed accordingly to industry standards and it is based on the determination of two specific signature peptides that report the single mAbs concentrations and on another one peptide, common to the two mAbs, that measures the total concentration of the two target proteins. It is shown that the total concentration is in agreement with the sum of the two measured single concentrations in spiked monkey serum samples. The consistency of the results will allow monitoring of the metabolic fate of different parts of the mAbs, at least in the central body compartment. This can then help to rationalize the design of the protein therapeutics modulating their stability accordingly.

Research paper thumbnail of Antiretrovirals: Simultaneous Determination of Five Protease Inhibitors and Three Nonnucleoside Transcriptase Inhibitors in Human Plasma by a Rapid High-Performance Liquid Chromatography–Mass Spectrometry Assay

Therapeutic Drug Monitoring, 2001

An analytical technique using liquid chromatography (LC) coupled with electrospray-mass spectrome... more An analytical technique using liquid chromatography (LC) coupled with electrospray-mass spectrometry (ESI--MS) has been developed for the simultaneous determination of five protease inhibitors (PIs): saquinavir, indinavir, ritonavir, nelfinavir, and amprenavir; and three non-nucleoside reverse transcriptase inhibitors (NNRTIs): nevirapine, delavirdine, and efavirenz, in human plasma. This assay allows the elution and identification of these drugs in a single run (10 minutes) using a linear gradient with water and acetonitrile. The procedure involves liquid--liquid extraction. High-performance liquid chromatography (HPLC) separation was achieved on a C18 reversed-phase column, with a linear gradient elution followed by mass spectrometry detection. The calibration curves, obtained by automatic process peak area integration, show a good linearity in a range of concentrations between 20 and 10,000 ng/mL (40--10,000 ng/mL for efavirenz). The limit of detection was approximately 10 ng/mL for seven drugs (25 ng/mL for efavirenz). The coefficients of variation (CV) were always less than 15% for both intraday and interday precision for each compound. The recovery of the eight drugs ranged from 88.5% to 100%. This novel LC/ESI--MS assay provides an excellent method for simultaneous quantitative monitoring of different components of the highly active antiretroviral treatments (HAARTs) in patients treated simultaneously with PIs and NNRTIs, and it has been successfully applied to therapeutic drug monitoring and pharmacokinetic studies.

Research paper thumbnail of A derivative of the glycopeptide A40926 produced by inactivation of the β-hydroxylase gene in Nonomuraea sp. ATCC39727: Derivative of the glycopeptide A40926

Fems Microbiol Lett, 2006

The actinomycete Nonomuraea sp. ATCC39727 produces the glycopeptide A40926.

Research paper thumbnail of PDF Planosporicin

Research paper thumbnail of Factors influencing cell fatty acid composition and A40926 antibiotic complex production in Nonomuraea sp. ATCC 39727

Journal of Industrial Microbiology & Biotechnology, 2008

A40926 is a glycopeptide antibiotic complex consisting of several structurally related factors. I... more A40926 is a glycopeptide antibiotic complex consisting of several structurally related factors. It is produced by fermentation of Nonomuraea sp. ATCC 39727 and the complex components differ in the structure of the fatty acid moiety linked to the aminoglucuronic acid unit. In previous work, we observed that the production of single factors in glycopeptide antibiotic complexes could be selectively enhanced by the addition of suitable precursors to the culture medium. In this contribution, we examine the effects of branched amino acid addition to fermentation of Nonomuraea sp. in a chemically defined minimal medium. The changes in the composition of cell fatty acids correlate to the fatty acid distribution within the A40926 complex in diverse cultivation conditions. Nonomuraea sp. prefers isobutyric, butyric and propionic acids as initiators of fatty acid biosynthesis. The relative amount of the produced fatty acids is significantly influenced by the availability of intermediates or final products from the amino acid catabolic pathways. Antibiotic complex composition closely reflects the cell fatty acid pattern, in agreement with the assumption that the antibiotic fatty acid moieties are synthesized by shortening the chain of cell fatty acids.

Research paper thumbnail of Valine Influences Production and Complex Composition of Glycopeptide Antibiotic A40926 in Fermentations of Nonomuraea sp. ATCC 39727

The Journal of Antibiotics, 2004

In actinomycetes the catabolism products of branched chain amino acids provide biosynthetic precu... more In actinomycetes the catabolism products of branched chain amino acids provide biosynthetic precursors for the formation of several lipid-containing antibiotics. We have determined in Nonomuraea sp. ATCC 39727 the effect of valine on production of glycopeptide antibiotic A40926, which is a complex of factors structurally differing in fatty acid moieties. Addition of valine to minimal medium increased A40926 production and modified complex composition towards a mono-component. Similar results were also obtained in a rich production medium.

Research paper thumbnail of A derivative of the glycopeptide A40926 produced by inactivation of the β-hydroxylase gene in Nonomuraea sp. ATCC39727

FEMS Microbiology Letters, 2000

The actinomycete Nonomuraea sp. ATCC39727 produces the glycopeptide A40926.

Research paper thumbnail of Novel Tetrapeptide Inhibitors of Bacterial Protein Synthesis Produced by a Streptomyces sp. †

Biochemistry, 2006

In the course of a microbial product screening aimed at the discovery of novel antibiotics acting... more In the course of a microbial product screening aimed at the discovery of novel antibiotics acting on bacterial protein synthesis, a complex of three structurally related tetrapeptides, namely, GE81112 factors A, B, and B1, was isolated from a Streptomyces sp. The screening was based on a cell-free assay of bacterial protein synthesis driven by a model mRNA containing natural initiation signals. In this study we report the production, isolation, and structure determination of these novel, potent and selective inhibitors of cell-free bacterial protein synthesis, which stably bind the 30S ribosomal subunit and inhibit the formation of fMet-puromycin. They did not inhibit translation by yeast ribosomes in vitro. Spectroscopic analyses revealed that they are tetrapeptides constituted by uncommon amino acids. While GE81112 factors A, B, and B1 were effective in inhibiting bacterial protein synthesis in vitro, they were less active against Gram-positive and Gram-negative bacterial cells. Cells grown in minimal medium were more susceptible to the compounds than those grown in rich medium, and this is most likely due to competition or regulation by medium components during peptide uptake. The novelty of the chemical structure and of the specific mode of action on the initiation phase of bacterial protein synthesis makes GE81112 a unique scaffold for designing new drugs.

Research paper thumbnail of A novel lantibiotic acting on bacterial cell wall synthesis produced by the uncommon actinomycete Planomonospora sp

Biochemistry, Jan 22, 2007

Important classes of antibiotics acting on bacterial cell wall biosynthesis, such as beta-lactams... more Important classes of antibiotics acting on bacterial cell wall biosynthesis, such as beta-lactams and glycopeptides, are used extensively in therapy and are now faced with a challenge because of the progressive spread of resistant pathogens. A discovery program was devised to target novel peptidoglycan biosynthesis inhibitors capable of overcoming these resistance mechanisms. The microbial products were first screened according to their differential activity against Staphylococcus aureus and its L-form. Then, activities insensitive to the addition of a beta-lactamase cocktail or d-alanyl-d-alanine affinity resin were selected. Thirty-five lantibiotics were identified from a library of broth extracts produced by 40,000 uncommon actinomycetes. Five of them showed structural characteristics that did not match with any known microbial metabolite. In this study, we report on the production, structure determination, and biological activity of one of these novel lantibiotics, namely, plano...

Research paper thumbnail of LC-MS Method for the Quantitation of Two Monoclonal Antibodies by Multiple Signature Peptides in Monkey Serum

Journal of Analytical & Bioanalytical Techniques, 2015

Evaluation of the in-vivo concentration of monoclonal antibody (mAb) mixtures is a challenging ta... more Evaluation of the in-vivo concentration of monoclonal antibody (mAb) mixtures is a challenging task. Here we report the application of an LC-MS bioanalytical method to quantify in monkey serum the Sym004, an equimolar mixture of two monoclonal antibodies, 992 mAb and 1024 mAb. This method has been assessed accordingly to industry standards and it is based on the determination of two specific signature peptides that report the single mAbs concentrations and on another one peptide, common to the two mAbs, that measures the total concentration of the two target proteins. It is shown that the total concentration is in agreement with the sum of the two measured single concentrations in spiked monkey serum samples. The consistency of the results will allow monitoring of the metabolic fate of different parts of the mAbs, at least in the central body compartment. This can then help to rationalize the design of the protein therapeutics modulating their stability accordingly.

Research paper thumbnail of Antiretrovirals: Simultaneous Determination of Five Protease Inhibitors and Three Nonnucleoside Transcriptase Inhibitors in Human Plasma by a Rapid High-Performance Liquid Chromatography–Mass Spectrometry Assay

Therapeutic Drug Monitoring, 2001

An analytical technique using liquid chromatography (LC) coupled with electrospray-mass spectrome... more An analytical technique using liquid chromatography (LC) coupled with electrospray-mass spectrometry (ESI--MS) has been developed for the simultaneous determination of five protease inhibitors (PIs): saquinavir, indinavir, ritonavir, nelfinavir, and amprenavir; and three non-nucleoside reverse transcriptase inhibitors (NNRTIs): nevirapine, delavirdine, and efavirenz, in human plasma. This assay allows the elution and identification of these drugs in a single run (10 minutes) using a linear gradient with water and acetonitrile. The procedure involves liquid--liquid extraction. High-performance liquid chromatography (HPLC) separation was achieved on a C18 reversed-phase column, with a linear gradient elution followed by mass spectrometry detection. The calibration curves, obtained by automatic process peak area integration, show a good linearity in a range of concentrations between 20 and 10,000 ng/mL (40--10,000 ng/mL for efavirenz). The limit of detection was approximately 10 ng/mL for seven drugs (25 ng/mL for efavirenz). The coefficients of variation (CV) were always less than 15% for both intraday and interday precision for each compound. The recovery of the eight drugs ranged from 88.5% to 100%. This novel LC/ESI--MS assay provides an excellent method for simultaneous quantitative monitoring of different components of the highly active antiretroviral treatments (HAARTs) in patients treated simultaneously with PIs and NNRTIs, and it has been successfully applied to therapeutic drug monitoring and pharmacokinetic studies.

Research paper thumbnail of A derivative of the glycopeptide A40926 produced by inactivation of the β-hydroxylase gene in Nonomuraea sp. ATCC39727: Derivative of the glycopeptide A40926

Fems Microbiol Lett, 2006

The actinomycete Nonomuraea sp. ATCC39727 produces the glycopeptide A40926.

Research paper thumbnail of PDF Planosporicin

Research paper thumbnail of Factors influencing cell fatty acid composition and A40926 antibiotic complex production in Nonomuraea sp. ATCC 39727

Journal of Industrial Microbiology & Biotechnology, 2008

A40926 is a glycopeptide antibiotic complex consisting of several structurally related factors. I... more A40926 is a glycopeptide antibiotic complex consisting of several structurally related factors. It is produced by fermentation of Nonomuraea sp. ATCC 39727 and the complex components differ in the structure of the fatty acid moiety linked to the aminoglucuronic acid unit. In previous work, we observed that the production of single factors in glycopeptide antibiotic complexes could be selectively enhanced by the addition of suitable precursors to the culture medium. In this contribution, we examine the effects of branched amino acid addition to fermentation of Nonomuraea sp. in a chemically defined minimal medium. The changes in the composition of cell fatty acids correlate to the fatty acid distribution within the A40926 complex in diverse cultivation conditions. Nonomuraea sp. prefers isobutyric, butyric and propionic acids as initiators of fatty acid biosynthesis. The relative amount of the produced fatty acids is significantly influenced by the availability of intermediates or final products from the amino acid catabolic pathways. Antibiotic complex composition closely reflects the cell fatty acid pattern, in agreement with the assumption that the antibiotic fatty acid moieties are synthesized by shortening the chain of cell fatty acids.

Research paper thumbnail of Valine Influences Production and Complex Composition of Glycopeptide Antibiotic A40926 in Fermentations of Nonomuraea sp. ATCC 39727

The Journal of Antibiotics, 2004

In actinomycetes the catabolism products of branched chain amino acids provide biosynthetic precu... more In actinomycetes the catabolism products of branched chain amino acids provide biosynthetic precursors for the formation of several lipid-containing antibiotics. We have determined in Nonomuraea sp. ATCC 39727 the effect of valine on production of glycopeptide antibiotic A40926, which is a complex of factors structurally differing in fatty acid moieties. Addition of valine to minimal medium increased A40926 production and modified complex composition towards a mono-component. Similar results were also obtained in a rich production medium.

Research paper thumbnail of A derivative of the glycopeptide A40926 produced by inactivation of the β-hydroxylase gene in Nonomuraea sp. ATCC39727

FEMS Microbiology Letters, 2000

The actinomycete Nonomuraea sp. ATCC39727 produces the glycopeptide A40926.

Research paper thumbnail of Novel Tetrapeptide Inhibitors of Bacterial Protein Synthesis Produced by a Streptomyces sp. †

Biochemistry, 2006

In the course of a microbial product screening aimed at the discovery of novel antibiotics acting... more In the course of a microbial product screening aimed at the discovery of novel antibiotics acting on bacterial protein synthesis, a complex of three structurally related tetrapeptides, namely, GE81112 factors A, B, and B1, was isolated from a Streptomyces sp. The screening was based on a cell-free assay of bacterial protein synthesis driven by a model mRNA containing natural initiation signals. In this study we report the production, isolation, and structure determination of these novel, potent and selective inhibitors of cell-free bacterial protein synthesis, which stably bind the 30S ribosomal subunit and inhibit the formation of fMet-puromycin. They did not inhibit translation by yeast ribosomes in vitro. Spectroscopic analyses revealed that they are tetrapeptides constituted by uncommon amino acids. While GE81112 factors A, B, and B1 were effective in inhibiting bacterial protein synthesis in vitro, they were less active against Gram-positive and Gram-negative bacterial cells. Cells grown in minimal medium were more susceptible to the compounds than those grown in rich medium, and this is most likely due to competition or regulation by medium components during peptide uptake. The novelty of the chemical structure and of the specific mode of action on the initiation phase of bacterial protein synthesis makes GE81112 a unique scaffold for designing new drugs.

Research paper thumbnail of A novel lantibiotic acting on bacterial cell wall synthesis produced by the uncommon actinomycete Planomonospora sp

Biochemistry, Jan 22, 2007

Important classes of antibiotics acting on bacterial cell wall biosynthesis, such as beta-lactams... more Important classes of antibiotics acting on bacterial cell wall biosynthesis, such as beta-lactams and glycopeptides, are used extensively in therapy and are now faced with a challenge because of the progressive spread of resistant pathogens. A discovery program was devised to target novel peptidoglycan biosynthesis inhibitors capable of overcoming these resistance mechanisms. The microbial products were first screened according to their differential activity against Staphylococcus aureus and its L-form. Then, activities insensitive to the addition of a beta-lactamase cocktail or d-alanyl-d-alanine affinity resin were selected. Thirty-five lantibiotics were identified from a library of broth extracts produced by 40,000 uncommon actinomycetes. Five of them showed structural characteristics that did not match with any known microbial metabolite. In this study, we report on the production, structure determination, and biological activity of one of these novel lantibiotics, namely, plano...