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首页> 外文期刊>Biotechnology Progress >Mass Spectrometric Analysis of Innovator,Counterfeit,and Follow-On Recombinant Human Growth Hormone
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Mass Spectrometric Analysis of Innovator,Counterfeit,and Follow-On Recombinant Human Growth Hormone

机译：创新者，假冒和后续重组人类生长激素的质谱分析

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We have performed a detailed characterization of recombinant human growth hormone that included the identification of the entire sequence with disulfide linkages as well as subtle modifications by a sensitive liquid chromatography coupled online with tandem mass spectrometry(LC-MS)approach using the accurate peptide mass(FTICR MS)and sequence assignment(MS/MS measurement).The extent of oxidation,deamidation,and chain cleavages were measured by the ratio of peak areas of the nonmodified peptide vs.the sum of peak area of the nonmodified and modified peptides in the same LC-MS analysis.The subtle but distinct differences were found in the recombinant human growth from the three manufacturers(the follow-on,counterfeit,and the original innovator products).In relative comparison,the follow-on product had the highest degree of oxidation at methionine residues,followed by the counterfeit product,and the original innovator product had the least amount of oxidation at all three sites with the similar oxidation order.In cases,the oxidation order was Metl4 > Metl25 > Metl70.In contrast,the follow-on had the least amount of deamidation at aspargine(Asnl49),and the counterfeit had the highest degree of deamidation at this site.For the chain cleavage,the follow-on product had the highest cleavage occurring at T 10 peptide(between Asn99 and Ser100),the counterfeit had the highest cleavage on T4 peptide,(between Glu30 and Phe31),and the original innovator product with the least amount of cleavages on both sites.These subtle but distinct differences are likely because of nonidentical manufacturing,formulation procedures,and storage conditions.
机译：我们已对重组人生长激素进行了详细的表征，包括通过具有精确肽质量的串联液相色谱与串联质谱（LC-MS）方法结合的灵敏液相色谱在线识别带有二硫键的整个序列以及进行细微修饰。 FTICR MS）和序列分配（MS / MS测量）。氧化，脱酰胺和链断裂的程度通过未修饰肽的峰面积比与未修饰肽中峰的总和来衡量。相同的LC-MS分析。在三个制造商（后续产品，假冒产品和原始创新产品）的重组人生长中发现了细微但明显的差异。相对而言，后续产品具有最高的程度甲硫氨酸残基的氧化反应，其次是假冒产品，而原始的创新产品在三个位置上的氧化程度最低类似的氧化顺序。在这种情况下，氧化顺序为Metl4> Metl25> Metl70。相比之下，后续分子在天冬酰胺（Asnl49）上的脱酰胺量最少，而假冒品在该部位的脱酰胺程度最高。链断裂，后续产物在T 10肽处断裂最高（在Asn99和Ser100之间），假冒产品在T4肽处断裂最高（在Glu30和Phe31之间），而原始的创新产品最少这两个细微但明显的差异可能是由于制造，配方程序和存储条件不同而引起的。

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