Stabilizing Peptide Fusion for Solving the Stability and Solubility Problems of Therapeutic Proteins

Lee, Eui Nam; Kim, Young Mok; Lee, Hye Ja; Park, Sang Woo; Jung, Han Young; Lee, Jae Myun; Ahn, Young Soo; Kim, Jongsun

doi:10.1007/s11095-005-6489-4

Cited by 50 publications

(54 citation statements)

References 48 publications

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“…The ATS-HTNV-N and ATS-SEOV-N fusion proteins appeared to be extremely heat-resistant, similar to other ATSfusion proteins reported previously (Lee et al, 2005;Park et al, 2002Park et al, , 2004. Therefore, they were easily purified to homogeneity by taking advantage of their heat resistance and by using conventional column chromatography techniques (Fig.…”

Section: Discussionsupporting

confidence: 72%

“…1B). Like other ATS fusion proteins (Lee et al, 2005;Park et al, 2002), ATS-HTNV-N and ATS-SEOV-N were extremely heat-resistant and there was no loss during heat treatment (data not shown). Taking advantage of the heat resistance, and using conventional column chromatography techniques, ATS-HTNV-N and ATS-SEOV-N (approximately 21 kDa each) were easily purified to homogeneity (Fig.…”

Section: Expression and Purification Of Ats-htnv-n And Ats-seov-n Fusmentioning

confidence: 86%

“…Previous studies have demonstrated that the introduction of a peptide derived from the C-terminal acidic tail of ␣-synuclein (ATS) into a heat-labile protein, GST, protects the fusion protein from heat-, pH-, and metal-induced protein aggregation (Lee et al, 2005;Park et al, 2002Park et al, , 2004. Previous studies have also shown that introduction of the ATS peptide significantly increases the stability and solubility of many therapeutic proteins (Lee et al, 2005). These results suggest that many biologically or medically important proteins that have solubility problems or stress-induced aggregation problems might be rendered more useful by introducing the ATS peptide.…”

Section: Introductionmentioning

confidence: 99%

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A soluble and heat-resistant form of hantavirus nucleocapsid protein for the serodiagnosis of HFRS

Park

Kim

2008

Journal of Virological Methods

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Section: Discussionsupporting

confidence: 72%

Section: Expression and Purification Of Ats-htnv-n And Ats-seov-n Fusmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A soluble and heat-resistant form of hantavirus nucleocapsid protein for the serodiagnosis of HFRS

Park

Kim

2008

Journal of Virological Methods

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“…In that case, it was speculated that structural issues or proteolytic degradation by carboxypeptidases was responsible [39][40][41]. Numerous methods have been proposed for protection of peptides and recombinant proteins from protease activity, including amidation or PEGylation of the C-terminus, acetylation of the amino (N)-terminus, and addition of protective amino acid motifs [42][43][44][45][46][47]. Two protective motifs, Gly-Gly and Pro-Pro-Ala, were assessed for their ability to enhance antibody responses to the ABKD vaccinogen.…”

Section: Discussionmentioning

confidence: 99%

Construction and analysis of variants of a polyvalent Lyme disease vaccine: Approaches for improving the immune response to chimeric vaccinogens

Earnhart

Marconi

2007

Vaccine

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There is currently no Lyme disease vaccine commercially available for use in humans. Outer surface protein C (OspC) of the Borrelia has been widely investigated as a potential vaccinogen. At least 38 OspC types have been defined. While the antibody response to OspC is protective, the range of protection is narrow due to the localization of protective epitopes within OspC type-specific domains. To develop a broadly protective vaccine, we previously constructed a tetravalent chimeric vaccinogen containing epitopes from OspC types A, B, K, and D. While this construct elicited bactericidal antibody against strains bearing each of the four OspC types, its solubility was low, and decreasing IgG titer to epitopes near the C-terminus of the construct was observed. In this report, construct solubility and immunogenicity were increased by dialysis against an Arg/Glu buffer. We also demonstrate the immunogenicity of the construct in alum. To further optimize epitope-specific immune responses, several constructs were generated with differing epitope organization or with putative C-terminal protective motifs. Analyses of murine antibody titers and isotype profiles induced by these constructs revealed that while the C-terminal tags did not enhance antibody titer, specific epitope reorganization and reiteration did. These analyses provide important information that can be exploited in the development of chimeric vaccinogens in general.

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“…A crucial aspect of every protein formulation is its stability under aggregate-favoring conditions, mainly heat and shaking (Lee et al, 2005). Here, restoration of activity after long incubation period was tested under both harsh and less detrimental condition.…”

Section: Discussionmentioning

confidence: 99%

Formation of multimeric antibodies for self-delivery of active monomers

Dekel

Machluf²,

Gefen

et al. 2017

Drug Delivery

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Proteins and peptides have been used as drugs for almost a century. Technological advances in the past 30 years have enabled the production of pure, stable proteins in vast amounts. In contrast, administration of proteins based on their native active conformation (and thus necessitating the use of subcutaneous injections) has remained solely unchanged. The therapeutic anti-HER2 humanized monoclonal immunoglobulin (IgG) Trastuzumab (Herceptin) is a first line of the treatment for breast cancer. Chicken IgY is a commercially important polyclonal antibody (Ab). These Abs were examined for their ability to self-assemble and form ordered aggregates, by several biophysical methods. Atomic force microscopy analyses revealed the formation of multimeric nanostructures. The biological activity of multimeric IgG or IgY particles was retained and restored, in a dilution/time-dependent manner. IgG activity was confirmed by a binding assay using HER2 + human breast cancer cell line, SKBR3, while IgY activity was confirmed by ELISA assay using the VP2 antigen. Competition assay with native Herceptin antibodies demonstrated that the binding availability of the multimer formulation remained unaffected. Under long incubation periods, IgG multimers retained five times more activity than native IgG. In conclusion, the multimeric antibody formulations can serve as a storage depositories and sustained-release particles. These two important characteristics make this formulation promising for future novel administration protocols and altogether bring to light a different conceptual approach for the future use of therapeutic proteins as self-delivery entities rather than conjugated/encapsulated to other bio-compounds.

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Stabilizing Peptide Fusion for Solving the Stability and Solubility Problems of Therapeutic Proteins

Abstract: Our data demonstrate that a more stable and soluble form of therapeutic proteins can be produced by fusing the ATS peptide.

Cited by 50 publications

References 48 publications

A soluble and heat-resistant form of hantavirus nucleocapsid protein for the serodiagnosis of HFRS

A soluble and heat-resistant form of hantavirus nucleocapsid protein for the serodiagnosis of HFRS

Construction and analysis of variants of a polyvalent Lyme disease vaccine: Approaches for improving the immune response to chimeric vaccinogens

Formation of multimeric antibodies for self-delivery of active monomers

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