Adamczyk Barbara, Albrecht Simone, Stöckmann Henning, Ghoneim Ibrahim M, Al-Eknah Marzook, Al-Busadah Khalid A S, Karlsson Niclas G, Carrington Stephen D, Rudd Pauline M
GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research and Training , Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland.
Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg , Gothenburg, 40530, Sweden.
J Proteome Res. 2016 Sep 2;15(9):3255-65. doi: 10.1021/acs.jproteome.6b00439. Epub 2016 Aug 3.
The dromedary camel (Camelus dromedarius) is an agriculturally important species of high economic value but of low reproductive efficiency. Serum and IgG N-glycosylation are affected by physiological and pathogenic changes and might therefore be a useful diagnostic tool in camel livestock management. This study presents the first comprehensive annotation of the N-glycome from dromedary camel serum as well as their single-domain and conventional antibodies and its subsequent application for camel pregnancy diagnostics. N-glycans were released by PNGaseF, labeled with 2-aminobenzamide (2-AB), and analyzed by hydrophilic interaction liquid chromatography with fluorescent detection (HILIC-UPLC-FLD), enzymatic sequencing and mass spectrometry (MS). The use of a high-throughput robotic platform for sample preparation allowed the rapid generation of glycomics data from pregnant (n = 8) and nonpregnant (n = 8) camels of the Majaheem and Wadha breed. IgG N-glycans dominate the glycan profile of camel serum and present a mixture of core-fucosylated and noncore-fucosylated N-glycans which can contain N-glycolylneuraminic and N-acetylneuraminic acid. Significant pregnancy-associated but breed-independent increases in galactosylation, core-fucosylation, sialylation, and decreases in serum O-acetylation were observed. The monitoring of IgG and serum N-glycosylation presents an attractive complementary test for camel pregnancy diagnostics and presents an interesting tool for biomarker discovery in camel health and breeding.
单峰骆驼(Camelus dromedarius)是一种具有重要农业意义、经济价值高但繁殖效率低的物种。血清和IgG的N-糖基化会受到生理和病理变化的影响,因此可能是骆驼畜牧管理中一种有用的诊断工具。本研究首次全面注释了单峰骆驼血清及其单域抗体和传统抗体的N-聚糖组,并将其随后应用于骆驼妊娠诊断。N-聚糖通过PNGaseF释放,用2-氨基苯甲酰胺(2-AB)标记,并通过亲水相互作用液相色谱-荧光检测(HILIC-UPLC-FLD)、酶促测序和质谱(MS)进行分析。使用高通量机器人平台进行样品制备,使得能够快速从Majaheem和Wadha品种的怀孕(n = 8)和未怀孕(n = 8)骆驼中生成糖组学数据。IgG N-聚糖在骆驼血清的聚糖谱中占主导地位,呈现出核心岩藻糖基化和非核心岩藻糖基化N-聚糖的混合物,其中可能含有N-羟乙酰神经氨酸和N-乙酰神经氨酸。观察到与妊娠相关但与品种无关的半乳糖基化、核心岩藻糖基化、唾液酸化显著增加,血清O-乙酰化减少。监测IgG和血清N-糖基化是骆驼妊娠诊断的一种有吸引力的补充检测方法,也是在骆驼健康和育种中发现生物标志物的一个有趣工具。