B***v
发帖数: 113 | 1 在一教科书上找到
“In addition to glycosylation of the two canonical Asn297 residues in the
Fc region, N-linked glycans are present in some 20 % of the Fab regions in
human IgG. These glycans include bi-antennary structures that are hyper-
galactosylated, ucosylated, and extensively sialylated.” |
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w******e
发帖数: 1187 | 2 想买n种biotinylated glycan，请教大人有没有大家常用的公司？多谢！ |
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B***v
发帖数: 113 | 3 多谢。
CDR上不会有N-glycan吧，附近有倒正常。
现在拿到个抗体vh/vl上有好几个，怀疑是半人半鼠或者半兔的杂交，因为人通常只有
一个我记得，不确定。 |
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m*********3
发帖数: 1425 | 4 在做Mab的glycosylation,有几个问题请教:
1.大家都知道Mab在Fc有个N-glycan,可我看文献，其实大约10-15%的Fab上也有N-
glycan,想请教一下大家对于这种有两个glycosylation的Mab怎么分析它的糖基化，现
在流行的用LC-MS,CE分析都是把糖酶切下来，然后label,但对于有两个糖基化的这个方
法似乎不算最好，对于有Fab N-glycan的大家一般怎么做？
2. 我看到有人用IdeS切Fab和Fc,比起Papain,哪个方法好一些呢？
3.另外对于Fc-fusion protein,还有O-glycan(比如Enbrel)，大家一般做glycopeptide
还chemically release呢？
4.还用过ETD确定glycosylation site,但ETD的效率很低，大家还有什么其它方法确定
glycosylation site吗？尤其是O-glycan, 谢谢。 |
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发帖数: 1 | 5 有了PRRA 基本可以排除实验室泄露可能被排除，可以转守围攻了
"There are also documented instances of the laboratory acquisition of SARS-
CoV-1 by laboratory personnel working under BSL-2 containment29,30. We must
therefore consider the possibility of a deliberate or inadvertent release of
SARS-CoV-2. In theory, it is possible that SARS-CoV-2 acquired the observed
RBD mutations site during adaptation to passage in cell culture, as has
been observed in studies with SARS-CoV5 as well as MERS-CoV31. However, the
acquisition of ... 阅读全帖 |
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r****o
发帖数: 105 | 6 本文献给YL
（十八）凝集素层析
我们用去垢剂溶解了膜蛋白之后，紧跟着的一步就是用Wheat Germ
Aglutinin(WGA) agarose来部分纯化insulin receptor。WGA是凝集素
(Lectin）中的一种，而Lectin指的是可以与糖基结合的蛋白质。现在常用
来做蛋白质纯化的凝集素绝大部分是从植物中提取出来的。绝大部分膜蛋
白的extracellular domain和分泌蛋白都有N-linked glycan 的糖基修饰。
根据这个性质，用固定有适当的凝集素的beads，比如WGA agarose或
者ConA agarose可以部分提纯膜蛋白。注意，只能是部分提纯，凝集素
层析并不能一步登天，让蛋白质比活力一下子提高上千倍，至多几十倍就
已经很不错了。但尽管如此，凝集素层析是一种很好的办法，因为buffer
条件非常温和，洗脱液是加了糖的buffer，透析什么的也方便，非常非常
适合于和其他层析方法偶连起来用。
纯化膜蛋白用的最主要的两种凝集素是WGA和ConA。谈到这里，
我先向大家介绍一下N-linked glycans。N |
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O***h
发帖数: 14 | 7 【 以下文字转载自 Biology 讨论区 】
发信人: OhYeh (comeon), 信区: Biology
标 题: Position available in boston
发信站: BBS 未名空间站 (Sun Apr 18 00:49:46 2010, 美东)
biopharm company in Boston looking for an experienced protein analyst,
qualifications: Ph.D or master with industrial experience in HPLC method
development,RP, SEC, MALs, glycan analysis, ELISA, IEF, Biacore. US work
permit a must.
Pls contact me offline: g****[email protected].
Thanks |
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r****o
发帖数: 105 | 8 My lab happens to study one important type of O-glycosylation.
O-glycosylations are carbohydrates linked through hydroxyl groups of
serine or threonine of the protein. There are different types of
O-glycosylations. They have different functions.
(1)O-GalNAc(mucin type)
Originally, O-glycosylation only refers to this type. This type
of glycosylation has usually extended chains(but less branched than
N-glycans). The first carbohydrate moiety linking to the protein is
N-acetylgalactosa |
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w******e
发帖数: 1187 | 9 首先得能发现个体的特异性差别。DNA/RNA的估计能指望NGS，protein要指望antibody
array/MS似乎还有很长一段路（希望aptamer array能迎头赶上呵呵）。小分子（
glycan
，metabolite，etc。最近出了个sarcosine）呢？lectin特异性太烂。希望aptamer
能顶上？不然也许只能指望HPLC之类能high throughput了？
其次得能把差别和各种physiological/pathological condition联系起来。这个ms进
展也太太太太慢了吧？就说cancer吧，有几个大家都服气的diagnostic/prognostic
/endpoint biomarker？一个PSA用了这么久，还一直被批。
最后要能根据这些差别来调节therapeutic regimen。Y/N型的或差别大的好办，学
herceptin
附赠个kit就是。检测分辨率不够的话呢？最好还是能够下一样的药，对不同的人达
到不同的效果，controlled release？也许就要在delivery method上做文章了？各
种deliv |
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j********e
发帖数: 52 | 10 Probing native protein structures by chemical cross-linking, mass
spectrometry and bioinformatics
Alexander Leitner, Thomas Walzthoeni, Abdullah Kahraman, Franz Herzog,
Oliver Rinner, Martin Beck, and Ruedi Aebersold
Mol Cell Proteomics published 31 March 2010, 10.1074/mcp.R000001-MCP201
Simultaneous glycan-peptide characterization using hydrophilic interaction
chromatography and parallel fragmentation by CID, HCD and ETD-MS applied to
the N-linked glycoproteome of Campylobacter jejuni
Nichollas |
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O***h
发帖数: 14 | 11 biopharm company in Boston looking for an experienced protein analyst,
qualifications: Ph.D or master with industrial experience in HPLC method
development,RP, SEC, MALs, glycan analysis, ELISA, IEF, Biacore. US work
permit a must.
Pls contact me offline: g****[email protected].
Thanks |
|
w******e
发帖数: 1187 | 12 糖是指glycan吗？请教jamie marth在这个领域的地位如何？多谢 |
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w******e
发帖数: 1187 | 13 any protein... aptamers can target any protein, glycan, drug
molecule, even metal ions:) |
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b******n
发帖数: 4225 | 14 http://www.mcponline.org/content/10/2/M000031-MCP201.abstract
Simultaneous Glycan-Peptide Characterization Using Hydrophilic Interaction
Chromatography and Parallel Fragmentation by CID, Higher Energy Collisional
Dissociation, and Electron Transfer Dissociation MS Applied to the N-Linked
Glycoproteome of Campylobacter jejuni
Nichollas E. Scott et al
Mol. Cell. Proteomics. 2011; 10:M000031-MCP201.
Email: s****[email protected]
Thanks a lot! |
|
E****A
发帖数: 184 | 15 http://www.ncbi.nlm.nih.gov/pubmed/21525513
Cold Spring Harb Perspect Biol. 2011 Apr 27 [Epub ahead of print]
Evolutionary Forces Shaping the Golgi Glycosylation Machinery: Why Cell
Surface
Glycans Are Universal to Living Cells.
Varki A.
Please send it to e****[email protected]
Thanks in advance! |
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l**********1
发帖数: 5204 | 16 大鼠/欧洲野兔/水䶄/鸡 的MHC 是越外(性)交: 路边的野花 不采白不采) 好似后代越advantageous哦
欧洲野兔 MHC paper:
Homozygosity at a class II MHC locus depresses female
reproductive ability in European brown hares
PubMed link:
//www.ncbi.nlm.nih.gov/pubmed/20731776
Wiki:
//en.wikipedia.org/wiki/European_Hare
水䶄MHC paper:
Selection Maintains MHC Diversity through a Natural Population Bottleneck
//www.ncbi.nlm.nih.gov/pubmed/22323362
Wiki:
//en.wikipedia.org/wiki/European_Water_Vole
据说恐龙演化过来的鸟类中的鸡 MHC paper
PNAS (2011)
Th... 阅读全帖 |
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w******e
发帖数: 1437 | 17 【 以下文字转载自 Chemistry 讨论区 】
发信人: wyysmile (wyy), 信区: Chemistry
标 题: paper help
发信站: BBS 未名空间站 (Fri Oct 25 21:48:00 2013, 美东)
I need the chapter 1 ( page 1-28 )in PDF format.
Chapter 1 : “De Novo Approaches to Monosaccharides and Complex Glycans”
Authors: Michael F. Cuccarese, Jiazhen J. Li, George A. O'Doherty
DOI: 10.1002/9783527658947.ch1
Book title: “Modern Synthetic Methods in Carbohydrate Chemistry: From
Monosaccharides to Complex Glycoconjugates”
Editors: Daniel B. Werz, Sébastien Vidal
非常感... 阅读全帖 |
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f*******K
发帖数: 34 | 18 用于常规蛋白质组研究，QE HF的performance已经完全可以和fusion媲美了。fusion的
优势在于选择灵活性和MS3 or 4用于修饰基团分析，比如glycans等，我们实验室
Fusion主要用于method development |
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q******g
发帖数: 3858 | 19 A red meat-derived glycan promotes inflammation and
cancer progression
A well known, epidemiologically reproducible risk factor for human
carcinomas is the long-term consumption of “red meat” of mammalian
origin. Although multiple theories have attempted to explain
this human-specific association, none have been conclusively
proven. We used an improved method to survey common foods
for free and glycosidically bound forms of the nonhuman sialic acid
N-glycolylneuraminic acid (Neu5Gc), showing tha... 阅读全帖 |
|
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B***v
发帖数: 113 | 21 human IgG 上应该是一个在Fc上，有多于一个的情况吗？
mouse IgG 是什么情况？
一时找不到答案，谢谢 |
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H****s
发帖数: 301 | 22 Case by case, it really depends on the germline/framework usage, VH/VL
pairing and CDR sequences. |
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d***s
发帖数: 1062 | 23 借此帖问问，抗体研发，都有些什么platform？phage display，ribosome display用
的多吗？ |
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H****s
发帖数: 301 | 24 用的最多的还是杂交瘤。对于in vitro display，每个公司的平台不太一样（有很大的
专利保护的因素在内）。如果你想要找这方面的工作的话，我建议你去开下这些会。一
是PEGS Summit，每年春天在波士顿开的；二是IBC抗体工程年会，每年12月初在San
Diego或者附近。多认识人，会很管用。现在in vitro display的位置非常少，一般出
来位置也都是associate的位置，对于有博士学位的人很不利。即使出来少数几个位置
，也都是从in vitro display的大牛实验室招。 |
|
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v**********m
发帖数: 5516 | 26 糖基化是在ER或高尔基体中完成，蛋白在这个成熟过程中不是完全折叠，所以所有可能
的氨基酸序列都可能被糖基化。宿主细胞的类型也很重要，同样的序列在不同的表达体
系中的糖修饰类型和修饰比例都会不同。 |
|
B***v
发帖数: 113 | 27 所以所有可能的氨基酸序列都可能被糖基化？
难道不是NxS/T motif？ |
|
|
U*****e
发帖数: 40 | 29 有不同的Glycoforms，然后又有不同的Isomers，确实复杂。 |
|
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B***v
发帖数: 113 | 31 能讲一下哪些NxS/T能被glycosylated，哪些不能，遵循什么规律，还是说所有的NxS/T
都能被glycosylated？ |
|
O***h
发帖数: 14 | 32 【 以下文字转载自 Biology 讨论区 】
发信人: OhYeh (comeon), 信区: Biology
标 题: Position available in boston
发信站: BBS 未名空间站 (Sun Apr 18 00:49:46 2010, 美东)
biopharm company in Boston looking for an experienced protein analyst,
qualifications: Ph.D or master with industrial experience in HPLC method
development,RP, SEC, MALs, glycan analysis, ELISA, IEF, Biacore. US work
permit a must.
Pls contact me offline: g****[email protected].
Thanks |
|
R*******1
发帖数: 6 | 33 Ad Description for BS/MS Sr Associate Scientist
Responsibilities:
• Work within the Analytical Protein Chemistry area to provide
analytical support for novel peptide and protein therapeutics in pre-
clinical and clinical development.
• Primary responsibilities include the identification and
characterization of post-translational modifications in protein therapeutic
candidates, using state of the art separations technology and mass
spectrometry instrumentation, with an emphasis ... 阅读全帖 |
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v**c
发帖数: 80 | 34 Hi Rockets11,
I saw this post on Chemistry board and very interested in this opening.
I am currently a Ph.D. student in Analytical Chemistry at Georgia State
University and will graduate in this May.
I have experience in the separation of small drugs with LC-MS/MS and CE-MS.
I also have some experience in the solid-phase synthesis and purification of
peptides.
If you can give me a email, I would like to send you my resume.
Thanks.
therapeutic
glycan |
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m********l
发帖数: 491 | 35 Is this job in Biogen? If not, can I send my resume to your email address?
Thanks.
therapeutic
glycan |
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y***p
发帖数: 57 | 36 主要做mAb，intact，sequence，PTM，glycan这些，还做一些leachable/extractable |
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w******e
发帖数: 1437 | 37 I need the chapter 1 ( page 1-28 )in PDF format.
Chapter 1 : “De Novo Approaches to Monosaccharides and Complex Glycans”
Authors: Michael F. Cuccarese, Jiazhen J. Li, George A. O'Doherty
DOI: 10.1002/9783527658947.ch1
Book title: “Modern Synthetic Methods in Carbohydrate Chemistry: From
Monosaccharides to Complex Glycoconjugates”
Editors: Daniel B. Werz, Sébastien Vidal
非常感谢您的热心帮助！ |
|
N******o
发帖数: 25 | 38 To support biologics process development in a CMO. PhD + 2 year post doc in
protein MS. intact mass, peptide mapping, glycan experience plus. prefer
QTOF experience.
Relocation assistance. No sponsorship.
Thanks! |
|
R*******1
发帖数: 6 | 39 Ad Description for BS/MS Sr Associate Scientist
Responsibilities:
• Work within the Analytical Protein Chemistry area to provide
analytical support for novel peptide and protein therapeutics in pre-
clinical and clinical development.
• Primary responsibilities include the identification and
characterization of post-translational modifications in protein therapeutic
candidates, using state of the art separations technology and mass
spectrometry instrumentation, with an emphasis ... 阅读全帖 |
|
l****n
发帖数: 711 | 40 1. GluC 和AspN比较常用，pepsin,chymo的特异性比较差。coverage是看confident
level的，和你怎么搜关系非常大。看看semi-enzymatic加了没，glycan搜出来没
2.找找blind modification search的软件。。。类似mascot error tolerance search
。说实话能公开找到的都不怎么好用。一般公司内部会有自己的软件或解决方法的 |
|
l****n
发帖数: 711 | 41 1. GluC 和AspN比较常用，pepsin,chymo的特异性比较差。coverage是看confident
level的，和你怎么搜关系非常大。看看semi-enzymatic加了没，glycan搜出来没
2.找找blind modification search的软件。。。类似mascot error tolerance search
。说实话能公开找到的都不怎么好用。一般公司内部会有自己的软件或解决方法的 |
|
h*****t
发帖数: 1226 | 42 Not about analysis, but if Fab has N-link glycan, if it is not required
for activity,try to mutant the sequence and remove it . It will help you
in the future development.
glycopeptide |
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m*********3
发帖数: 1425 | 43 Sometimes, Fab N-glycan is important for activity. Indeed, Cetuximab has two
N-glycosylation sites. Furthermore, other glycoprotein such as EPO has
three N-glycosylation and one O-glycosylation, you can't just mutate the
sequence and remove all of them. |
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b****u
发帖数: 2771 | 44
glycopeptide
做o-glycan site ID 用CID不一定不行,你用啥仪器做 |
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o**s
发帖数: 891 | 45 看来药厂contractor不是一个上佳的选择，我也隐约有这种感觉。我也知道口头上说将
来如何如何其实都不能指望太多，只是觉得只要工作内容中有好的东西，那就也可以考
虑。
回到现在的工作，虽说是个postdoc,却也因为有很多难得的条件，所以才有些犹豫。不
知道自己过多的考虑技术方向是不是幼稚了。因为也听人说，进了工业界以后，增加的
不仅是业界的经验，也会更多的接触工业界的人脉关系和机会，这些是在学术界难以相
比的。所以进入工业界的第一步最难，有机会就应该好好珍惜，后面再跳槽相对容易多
了。
当然，我知道这个东西很难一概而论的回答，它取决于每个人的性格，和具体的工作领
域。
我并不是一个特别内向的人，爱交朋友，但是却特别不爱开口求人帮忙。 经常是不到
万不得已实在不行才求人的。现在正在试图慢慢改进。但从前找工作都是靠一份份投简
历的。所以很珍惜这个offer.
另一方面，我之所以觉得现在这个offer不是我很理想的方向，是因为发现主要是做mrm
quantification.我个人觉得这个技术要求比较不高，工业界需求机会也并不是很多。
似乎做蛋白质或者抗体PTM, glycan什么的变... 阅读全帖 |
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a****o
发帖数: 1786 | 46 【 以下文字转载自 Biology 讨论区 】
发信人: OhYeh (comeon), 信区: Biology
标 题: Position available in boston
发信站: BBS 未名空间站 (Sun Apr 18 00:49:46 2010, 美东)
biopharm company in Boston looking for an experienced protein analyst,
qualifications: Ph.D or master with industrial experience in HPLC method
development,RP, SEC, MALs, glycan analysis, ELISA, IEF, Biacore. US work
permit a must.
Pls contact me offline: g****[email protected].
Thanks |
|
