Quality screening of incorrectly folded soluble aggregates from functional recombinant proteins

Soon Bin Kwon, Ji Eun Yu, Jihoon Kim, Hana Oh, Chan Park, Jinhee Lee, Baik Lin Seong

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Solubility is the prime criterion for determining the quality of recombinant proteins, yet it often fails to represent functional activity due to the involvement of non-functional, misfolded, soluble aggregates, which compromise the quality of recombinant proteins. However, guidelines for the quality assessment of soluble proteins have neither been proposed nor rigorously validated experimentally. Using the aggregation-prone enhanced green-fluorescent protein (EGFP) folding reporter system, we evaluated the folding status of recombinant proteins by employing the commonly used sonication and mild lysis of recombinant host cells. We showed that the differential screening of solubility and folding competence is crucial for improving the quality of recombinant proteins without sacrificing their yield. These results highlight the importance of screening out incorrectly folded soluble aggregates at the initial purification step to ensure the functional quality of recombinant proteins.

Original languageEnglish
Article number907
JournalInternational journal of molecular sciences
Volume20
Issue number4
DOIs
Publication statusPublished - 2019 Feb 2

Fingerprint

Recombinant proteins
Recombinant Proteins
Screening
screening
proteins
folding
Solubility
Protein folding
Sonication
Protein Folding
solubility
Mental Competency
Purification
Agglomeration
Guidelines
purification
Proteins
cells

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

Cite this

Kwon, Soon Bin ; Yu, Ji Eun ; Kim, Jihoon ; Oh, Hana ; Park, Chan ; Lee, Jinhee ; Seong, Baik Lin. / Quality screening of incorrectly folded soluble aggregates from functional recombinant proteins. In: International journal of molecular sciences. 2019 ; Vol. 20, No. 4.
@article{137885eee6d649f1b689327fd6805caf,
title = "Quality screening of incorrectly folded soluble aggregates from functional recombinant proteins",
abstract = "Solubility is the prime criterion for determining the quality of recombinant proteins, yet it often fails to represent functional activity due to the involvement of non-functional, misfolded, soluble aggregates, which compromise the quality of recombinant proteins. However, guidelines for the quality assessment of soluble proteins have neither been proposed nor rigorously validated experimentally. Using the aggregation-prone enhanced green-fluorescent protein (EGFP) folding reporter system, we evaluated the folding status of recombinant proteins by employing the commonly used sonication and mild lysis of recombinant host cells. We showed that the differential screening of solubility and folding competence is crucial for improving the quality of recombinant proteins without sacrificing their yield. These results highlight the importance of screening out incorrectly folded soluble aggregates at the initial purification step to ensure the functional quality of recombinant proteins.",
author = "Kwon, {Soon Bin} and Yu, {Ji Eun} and Jihoon Kim and Hana Oh and Chan Park and Jinhee Lee and Seong, {Baik Lin}",
year = "2019",
month = "2",
day = "2",
doi = "10.3390/ijms20040907",
language = "English",
volume = "20",
journal = "International Journal of Molecular Sciences",
issn = "1661-6596",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "4",

}

Quality screening of incorrectly folded soluble aggregates from functional recombinant proteins. / Kwon, Soon Bin; Yu, Ji Eun; Kim, Jihoon; Oh, Hana; Park, Chan; Lee, Jinhee; Seong, Baik Lin.

In: International journal of molecular sciences, Vol. 20, No. 4, 907, 02.02.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Quality screening of incorrectly folded soluble aggregates from functional recombinant proteins

AU - Kwon, Soon Bin

AU - Yu, Ji Eun

AU - Kim, Jihoon

AU - Oh, Hana

AU - Park, Chan

AU - Lee, Jinhee

AU - Seong, Baik Lin

PY - 2019/2/2

Y1 - 2019/2/2

N2 - Solubility is the prime criterion for determining the quality of recombinant proteins, yet it often fails to represent functional activity due to the involvement of non-functional, misfolded, soluble aggregates, which compromise the quality of recombinant proteins. However, guidelines for the quality assessment of soluble proteins have neither been proposed nor rigorously validated experimentally. Using the aggregation-prone enhanced green-fluorescent protein (EGFP) folding reporter system, we evaluated the folding status of recombinant proteins by employing the commonly used sonication and mild lysis of recombinant host cells. We showed that the differential screening of solubility and folding competence is crucial for improving the quality of recombinant proteins without sacrificing their yield. These results highlight the importance of screening out incorrectly folded soluble aggregates at the initial purification step to ensure the functional quality of recombinant proteins.

AB - Solubility is the prime criterion for determining the quality of recombinant proteins, yet it often fails to represent functional activity due to the involvement of non-functional, misfolded, soluble aggregates, which compromise the quality of recombinant proteins. However, guidelines for the quality assessment of soluble proteins have neither been proposed nor rigorously validated experimentally. Using the aggregation-prone enhanced green-fluorescent protein (EGFP) folding reporter system, we evaluated the folding status of recombinant proteins by employing the commonly used sonication and mild lysis of recombinant host cells. We showed that the differential screening of solubility and folding competence is crucial for improving the quality of recombinant proteins without sacrificing their yield. These results highlight the importance of screening out incorrectly folded soluble aggregates at the initial purification step to ensure the functional quality of recombinant proteins.

UR - http://www.scopus.com/inward/record.url?scp=85061984862&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85061984862&partnerID=8YFLogxK

U2 - 10.3390/ijms20040907

DO - 10.3390/ijms20040907

M3 - Article

VL - 20

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1661-6596

IS - 4

M1 - 907

ER -