Targeted serum glycoproteomics for the discovery of lung cancer‐associated glycosylation disorders using lectin‐coupled ProteinChip arrays
Targeted serum glycoproteomics for the discovery of lung cancer‐associated glycosylation disorders using lectin‐coupled ProteinChip arrays
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DOI：
10.1002/pmic.200800374
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发表时间：
2009-04
期刊：
影响因子：
3.4
通讯作者：
K. Ueda;Y. Fukase;T. Katagiri;N. Ishikawa;S. Irie;Takashi Sato;H. Ito;H. Nakayama;Y. Miyagi;E. Tsuchiya;N. Kohno;M. Shiwa;Yusuke Nakamura;Y. Daigo
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文献类型：
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作者：
K. Ueda;Y. Fukase;T. Katagiri;N. Ishikawa;S. Irie;Takashi Sato;H. Ito;H. Nakayama;Y. Miyagi;E. Tsuchiya;N. Kohno;M. Shiwa;Yusuke Nakamura;Y. Daigo
为了筛选癌症患者血清中显示异常唾液酸化模式的糖蛋白并将此类信息应用于生物标志物鉴定，我们使用从肺癌患者和对照个体获得的血清，结合凝集素偶联蛋白芯片阵列（Jacalin或SNA）进行了SELDI-TOF MS分析。我们的方法包括三个过程：（i）去除血清中14种丰富的蛋白质，（ii）用凝集素偶联的蛋白质芯片阵列富集糖蛋白，（iii）用酸性糖蛋白相容基质进行SELDI-TOF MS分析。我们使用Jacalin‐和SNA‐ProteinChips鉴定了41个蛋白质峰，显示癌症组和对照组之间的峰水平存在显著差异（p<0.05）。其中，我们鉴定了载脂蛋白C-III中Neu 5Ac（α 2，6）Gal/GalNAc结构的缺失，此外，使用另外一组60名肺腺癌患者和30名正常对照的后续验证实验证明，在α2缺失的情况下，血清apoC-III的频率更高，与对照相比，肺癌患者中的6-连接Neu 5Ac残基。我们的研究结果表明，凝集素偶联的蛋白质芯片技术可以高通量和特异性地识别癌症相关的异常糖基化，并暗示其适用于其他疾病的研究的可能性。
To screen for glycoproteins showing aberrant sialylation patterns in sera of cancer patients and apply such information for biomarker identification, we performed SELDI‐TOF MS analysis coupled with lectin‐coupled ProteinChip arrays (Jacalin or SNA) using sera obtained from lung cancer patients and control individuals. Our approach consisted of three processes (i) removal of 14 abundant proteins in serum, (ii) enrichment of glycoproteins with lectin‐coupled ProteinChip arrays, and (iii) SELDI‐TOF MS analysis with acidic glycoprotein‐compatible matrix. We identified 41 protein peaks showing significant differences (p<0.05) in the peak levels between the cancer and control groups using the Jacalin‐ and SNA‐ProteinChips. Among them, we identified loss of Neu5Ac (α2,6) Gal/GalNAc structure in apolipoprotein C‐III (apoC‐III) in cancer patients through subsequent MALDI‐QIT‐TOF MS/MS. Furthermore, subsequent validation experiments using an additional set of 60 lung adenocarcinoma patients and 30 normal controls demonstrated that there is a higher frequency of serum apoC‐III with loss of α2,6‐linkage Neu5Ac residues in lung cancer patients compared to controls. Our results have demonstrated that lectin‐coupled ProteinChip technology allows the high‐throughput and specific recognition of cancer‐associated aberrant glycosylations, and implied a possibility of its applicability to studies on other diseases.