여름정기학술대회
2022여름초록
발표자 및 발표 내용
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포스터발표 |
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Brief Oral Presentation 발표신청 |
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주저자
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국가 |
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공동저자
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접수자
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Keratan
sulfate (KS)-proteoglycan is a linear polymer consisting of repeating
disaccharide with a negative charge imparted by sulfate. KS is a sophisticated
molecule with a diverse structure, and unique functional roles in the cornea,
cartilage catabolism, and joint formation. In particular, accumulation of
undegraded KS by enzyme deficiency results in skeletal cartilage dysplasia and
lysosomal storage disorders such as mucopolysaccharidosis IVA and IVB.
Therefore, detection and quantitation of KS are important for the diagnosis and
treatment of related diseases, but it is still analytically challenging due to
low concentration of KS in biological samples and the difficulty of selective
separation of KS. In this study, we developed a highly sensitive and selective
method for quantitating KS in mouse urine and liver using the combination of
solid-phase extraction (SPE) and LC-MS/MS MRM. In addition, the Folch method
and spin column were used to efficiently remove liver lipids and urine
metabolites, respectively. Thereafter, KS was selectively fractionated by SPE
with a PGC cartridge according to its molecule size and polarity (acidity,
pKa). Quantitation using LC-MS/MS MRM were optimized and validated by polymer
KS, LacNAc(1S), LacNAc(2S) standards and Keratanase II prior to actual sample
analysis. In the case of urine, KS was quantified in the range of 1-20 pg/mL,
and in the liver, it was quantified in the range of 0.1-20 pg/mg.
Our new platform with high sensitivity and selectivity can be widely used for
diagnosis and treatment of KS-related disease.
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