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LINK . SPRINGER . COM {}

  1. Analyzed Page
  2. Matching Content Categories
  3. CMS
  4. Monthly Traffic Estimate
  5. How Does Link.springer.com Make Money
  6. Keywords
  7. Topics
  8. Questions
  9. Schema
  10. External Links
  11. Analytics And Tracking
  12. Libraries
  13. CDN Services

We are analyzing https://link.springer.com/article/10.1007/s12033-025-01450-4.

Title:
Toward the Development of a Biosimilar Variant of Glucarpidase (Carboxypeptidase G2): Secretory Production, Optimization, and Immobilization | Molecular Biotechnology
Description:
Carboxypeptidase G2 (CPG2) detoxifies high-dose methotrexate in cancer or autoimmune therapies. Additionally, CPG2 activates prodrugs at the tumor site in engineered CAR-T cell therapy. Since CPG2 cleaves glutamate from the substrate, it also has potential applications in enhancing food flavors and developing biosensors. However, cytoplasmic expression of recombinant CPG2 often leads to inclusion body formation, necessitating secretory production to improve product quality and streamline downstream industrial processes. This study aimed to perform a comprehensive in silico analysis to identify an optimal signal peptide for CPG2 secretion. The pelB leader sequence was selected for excretory production using the pET22b vector in Escherichia coli. The enzyme was successfully secreted into both the periplasmic space of bacterial cells and the culture medium with activities of about 0.038 UmL‒1 and 0.0285 UmL‒1, respectively. Optimal expression conditions were achieved at 25 °C with 0.5-mM IPTG, leading to enhanced yields. In the second step, we aimed to optimize the culture media composition. Therefore, a statistical design of experiments (DOE) approach in Modde software was conducted, evaluating eleven variables via a fractional factorial design (FFD). A central composite face (CCF) design was applied to establish an empirical model for maximizing periplasmic CPG2 production. The final aim was to immobilize the whole E. coli cells in calcium alginate beads for higher metabolic activity, improved growth rates, and plasmid stability. Engineered bacterial cells were entrapped in a mixture of sodium alginate and calcium chloride solution and incubated at 4 °C, 400 rpm for 60 min. Immobilization of CPG2-producing E. coli cells in 2% (w/v) calcium alginate beads yielded spherical beads, maintaining enzyme stability for 10 days, with peak activity on day 5. Conclusively, improved CPG2 solubility, facilitated protein purification, and enzyme stability over multiple reaction cycles offer a cost-effective solution for industrial purposes.
Website Age:
28 years and 1 months (reg. 1997-05-29).

Matching Content Categories {📚}

  • Education
  • Science
  • Health & Fitness

Content Management System {📝}

What CMS is link.springer.com built with?

Custom-built

No common CMS systems were detected on Link.springer.com, and no known web development framework was identified.

Traffic Estimate {📈}

What is the average monthly size of link.springer.com audience?

🌠 Phenomenal Traffic: 5M - 10M visitors per month


Based on our best estimate, this website will receive around 8,170,536 visitors per month in the current month.

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How Does Link.springer.com Make Money? {💸}

We see no obvious way the site makes money.

Not all websites are made for profit; some exist to inform or educate users. Or any other reason why people make websites. And this might be the case. Link.springer.com might be earning cash quietly, but we haven't detected the monetization method.

Keywords {🔍}

article, google, scholar, pubmed, cas, coli, central, production, protein, journal, biotechnology, escherichia, expression, signal, recombinant, engineering, research, immobilization, enzyme, microbiology, peptide, carboxypeptidase, cell, sciences, shiraz, hemmati, purification, peptides, optimization, bacterial, cells, nature, applications, proteins, supplementary, glucarpidase, cancer, biological, microbial, current, analysis, therapy, calcium, metabolic, chemistry, file, docx, molecular, ghasemi, secretion,

Topics {✒️}

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Questions {❓}

  • Can a signal sequence become too hydrophobic?
  • Is enzyme immobilization a mature discipline?

Schema {🗺️}

WebPage:
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         headline:Toward the Development of a Biosimilar Variant of Glucarpidase (Carboxypeptidase G2): Secretory Production, Optimization, and Immobilization
         description:Carboxypeptidase G2 (CPG2) detoxifies high-dose methotrexate in cancer or autoimmune therapies. Additionally, CPG2 activates prodrugs at the tumor site in engineered CAR-T cell therapy. Since CPG2 cleaves glutamate from the substrate, it also has potential applications in enhancing food flavors and developing biosensors. However, cytoplasmic expression of recombinant CPG2 often leads to inclusion body formation, necessitating secretory production to improve product quality and streamline downstream industrial processes. This study aimed to perform a comprehensive in silico analysis to identify an optimal signal peptide for CPG2 secretion. The pelB leader sequence was selected for excretory production using the pET22b vector in Escherichia coli. The enzyme was successfully secreted into both the periplasmic space of bacterial cells and the culture medium with activities of about 0.038 UmL‒1 and 0.0285 UmL‒1, respectively. Optimal expression conditions were achieved at 25 °C with 0.5-mM IPTG, leading to enhanced yields. In the second step, we aimed to optimize the culture media composition. Therefore, a statistical design of experiments (DOE) approach in Modde software was conducted, evaluating eleven variables via a fractional factorial design (FFD). A central composite face (CCF) design was applied to establish an empirical model for maximizing periplasmic CPG2 production. The final aim was to immobilize the whole E. coli cells in calcium alginate beads for higher metabolic activity, improved growth rates, and plasmid stability. Engineered bacterial cells were entrapped in a mixture of sodium alginate and calcium chloride solution and incubated at 4 °C, 400 rpm for 60 min. Immobilization of CPG2-producing E. coli cells in 2% (w/v) calcium alginate beads yielded spherical beads, maintaining enzyme stability for 10 days, with peak activity on day 5. Conclusively, improved CPG2 solubility, facilitated protein purification, and enzyme stability over multiple reaction cycles offer a cost-effective solution for industrial purposes.
         datePublished:2025-06-04T00:00:00Z
         dateModified:2025-06-04T00:00:00Z
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                     address:
                        name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
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      headline:Toward the Development of a Biosimilar Variant of Glucarpidase (Carboxypeptidase G2): Secretory Production, Optimization, and Immobilization
      description:Carboxypeptidase G2 (CPG2) detoxifies high-dose methotrexate in cancer or autoimmune therapies. Additionally, CPG2 activates prodrugs at the tumor site in engineered CAR-T cell therapy. Since CPG2 cleaves glutamate from the substrate, it also has potential applications in enhancing food flavors and developing biosensors. However, cytoplasmic expression of recombinant CPG2 often leads to inclusion body formation, necessitating secretory production to improve product quality and streamline downstream industrial processes. This study aimed to perform a comprehensive in silico analysis to identify an optimal signal peptide for CPG2 secretion. The pelB leader sequence was selected for excretory production using the pET22b vector in Escherichia coli. The enzyme was successfully secreted into both the periplasmic space of bacterial cells and the culture medium with activities of about 0.038 UmL‒1 and 0.0285 UmL‒1, respectively. Optimal expression conditions were achieved at 25 °C with 0.5-mM IPTG, leading to enhanced yields. In the second step, we aimed to optimize the culture media composition. Therefore, a statistical design of experiments (DOE) approach in Modde software was conducted, evaluating eleven variables via a fractional factorial design (FFD). A central composite face (CCF) design was applied to establish an empirical model for maximizing periplasmic CPG2 production. The final aim was to immobilize the whole E. coli cells in calcium alginate beads for higher metabolic activity, improved growth rates, and plasmid stability. Engineered bacterial cells were entrapped in a mixture of sodium alginate and calcium chloride solution and incubated at 4 °C, 400 rpm for 60 min. Immobilization of CPG2-producing E. coli cells in 2% (w/v) calcium alginate beads yielded spherical beads, maintaining enzyme stability for 10 days, with peak activity on day 5. Conclusively, improved CPG2 solubility, facilitated protein purification, and enzyme stability over multiple reaction cycles offer a cost-effective solution for industrial purposes.
      datePublished:2025-06-04T00:00:00Z
      dateModified:2025-06-04T00:00:00Z
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         Carboxypeptidase G2
         Glutamate
         Immobilization
         Methotrexate
         Periplasm
         Signal peptide
         Biotechnology
         Biochemistry
         general
         Cell Biology
         Protein Science
         Biological Techniques
         Human Genetics
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         name:Molecular Biotechnology
         issn:
            1559-0305
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         type:
            Periodical
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         name:Springer US
         logo:
            url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
            type:ImageObject
         type:Organization
      author:
            name:Fatemeh Moradbeygi
            affiliation:
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Mohammad Bagher Ghoshoon
            affiliation:
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Issa Sadeghian
            affiliation:
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Biotechnology Incubator, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Anahita Moradi
            affiliation:
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
                  name:Universitätsklinikum Freiburg
                  address:
                     name:Universitätsklinikum Freiburg, Freiburg, Germany
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Younes Ghasemi
            affiliation:
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Mohammad Hossein Morowvat
            affiliation:
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Shiva Hemmati
            url:http://orcid.org/0000-0001-9071-9569
            affiliation:
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
                  type:Organization
                  name:Shiraz University of Medical Sciences
                  address:
                     name:Biotechnology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
                     type:PostalAddress
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      name:Molecular Biotechnology
      issn:
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      name:Springer US
      logo:
         url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
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      name:Shiraz University of Medical Sciences
      address:
         name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Biotechnology Incubator, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Universitätsklinikum Freiburg
      address:
         name:Universitätsklinikum Freiburg, Freiburg, Germany
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
      name:Shiraz University of Medical Sciences
      address:
         name:Biotechnology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
         type:PostalAddress
ImageObject:
      url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
Person:
      name:Fatemeh Moradbeygi
      affiliation:
            name:Shiraz University of Medical Sciences
            address:
               name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
      name:Mohammad Bagher Ghoshoon
      affiliation:
            name:Shiraz University of Medical Sciences
            address:
               name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
      name:Issa Sadeghian
      affiliation:
            name:Shiraz University of Medical Sciences
            address:
               name:Biotechnology Incubator, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
      name:Anahita Moradi
      affiliation:
            name:Shiraz University of Medical Sciences
            address:
               name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
            name:Universitätsklinikum Freiburg
            address:
               name:Universitätsklinikum Freiburg, Freiburg, Germany
               type:PostalAddress
            type:Organization
      name:Younes Ghasemi
      affiliation:
            name:Shiraz University of Medical Sciences
            address:
               name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
            name:Shiraz University of Medical Sciences
            address:
               name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
      name:Mohammad Hossein Morowvat
      affiliation:
            name:Shiraz University of Medical Sciences
            address:
               name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
      name:Shiva Hemmati
      url:http://orcid.org/0000-0001-9071-9569
      affiliation:
            name:Shiraz University of Medical Sciences
            address:
               name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
            name:Shiraz University of Medical Sciences
            address:
               name:Biotechnology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
               type:PostalAddress
            type:Organization
      email:[email protected]
PostalAddress:
      name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Biotechnology Incubator, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Universitätsklinikum Freiburg, Freiburg, Germany
      name:Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
      name:Biotechnology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
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External Links {🔗}(469)

Analytics and Tracking {📊}

  • Google Tag Manager

Libraries {📚}

  • Clipboard.js
  • Prism.js

CDN Services {📦}

  • Crossref

4.47s.