Octet® 非標記分子互作分析系統(tǒng)采用實時的非標記分析技術(shù)，用于測定親和力、動力學和抗體/蛋白質(zhì)定量。這種無流路的儀器平臺基于生物層干涉（BLI）技術(shù)而設計，與傳統(tǒng)的非標記技術(shù)相比，具有眾多優(yōu)勢。

2024年上半年（截至6月30日在線發(fā)表）的國內(nèi)論文中，使用賽多利斯Octet® 數(shù)據(jù)的國內(nèi)論文呈現(xiàn)井噴狀態(tài)。陳老師統(tǒng)計了2024年上半年約200篇國內(nèi)論文（預計總數(shù)超過200篇），這些文章分布于130個單位， 詳細列表在文末哦~老規(guī)矩，讓我們先按照單位和應用進行統(tǒng)計。
 

圖1. 2024上半年Octet® 文章第一作者單位分布
 

圖2. 2024H1 Octet® 文章應用分布
 
中國藥科大學、北京協(xié)和醫(yī)學院、南京大學、復旦大學、浙江大學和中山大學等單位發(fā)表的文章均在4篇以上。其中，中國藥科大學論文數(shù)達到了10篇。這表明，Octet® 因其操作簡單、成本低、數(shù)據(jù)準確和儀器穩(wěn)定性好等優(yōu)點，可以幫助科學家快速取得科研成果。

從應用方向上看，依舊涉及到多個方向。其中小分子的應用超過蛋白和抗體應用的文章數(shù)量，牢牢占據(jù)應用方向榜首！BLI技術(shù)對溶劑（DMSO）不敏感，假陽性低，高靈敏度等優(yōu)點可以幫助研究人更快，更高質(zhì)量的獲得了小分子檢測的結(jié)果。


今天，我們就來挑選幾篇代表性文章來一一解讀：


浙江大學——細胞與分子互作^[1]
 
全球氣候變化伴隨著二氧化碳（CO₂）富集和高溫（HT）脅迫。研究發(fā)現(xiàn)，番茄外源質(zhì)外糖 （Glc）處理增強了番茄在環(huán)境CO₂條件下對HT脅迫的恢復力�；�于細胞的生物層干涉法（Octet® ）、亞細胞定位和分裂熒光素酶測定表明，Glc 與番茄 G 蛋白信號轉(zhuǎn)導 1 （RGS1） 調(diào)節(jié)因子結(jié)合并誘導 RGS1 內(nèi)吞作用，從而以濃度依賴性方式誘導 RGS1-G 蛋白α亞基 （GPA1） 解離，這是 CO₂-Glc 誘導的耐熱性升高所必需的。這些信息增強了對Glc-G蛋白信號傳導功能在應對全球氣候變化的脅迫恢復力中的理解，并將有助于開發(fā)植物恢復力的基因工程方法。
 

圖3. 將原生質(zhì)體固化在Octet® 傳感器上，結(jié)合各種單糖，發(fā)現(xiàn)表達RGS1的細胞主要結(jié)合葡萄糖，但是將RGS1突變后，與葡萄糖結(jié)合變?nèi)?br />  
Octet® 因為浸入即讀的方法，更適合檢測比較大的分析物，比較細胞。


西湖大學——血清抗體濃度測定^[2]
 
第三劑滅活新型冠狀病毒疫苗的抗體反應對疫苗的保護作用至關(guān)重要。西湖大學工學院對接種疫苗的15人的九個月里七個時間點進行了微量取樣和靜脈穿刺取樣，并建立了基于Octet® 的光纖生物層干涉法測量(FO-BLI)來測試血清中的中和抗體，可以在幾分鐘內(nèi)完成測試。研究結(jié)果揭示發(fā)現(xiàn)在第三次給藥后，野生型變體的血清NAb水平在7天后增加了2.9倍，在一個月內(nèi)增加了3.3倍，隨后下降，并在三個月后變得不可檢測。針對原始株與變異株的中和能力檢測, FO-BLI與假病毒中和試驗高度相關(guān)(Pearson 相關(guān)系數(shù)為0.983)。基于FO-BLI的濃度測試因為其快速，并且與功能性匹配，有潛力在疾病預防和疫苗開發(fā)中有著更大的應用。
 

圖4. A圖：Octet® 建立血清中和抗體的檢測方法，將受體ACE2結(jié)合在傳感器上，與HRP偶聯(lián)的不同病毒株的RBD和血清的混合物孵育，然后用底物放大信號；B圖：這種方法測試的標準抗體的靈敏度可以達到20ng/mL；
 

D圖：FO-BLI的檢測結(jié)果與病毒中和實驗PVNT（IC50）有良好的相關(guān)性

使用生物層干涉技術(shù)，可以快速建立濃度檢測方法。
 
 
浙江工業(yè)大學/北大醫(yī)學院——分子垂釣^[3]

研究者建立了一種基于Octet® 的垂釣技術(shù)，研究人員將GNAS蛋白固化至SA傳感器表面，再與中藥參芪降糖顆粒（SJG）提取物結(jié)合，隨后提取物被解離至洗脫液中，將洗脫液使用UHPLC-Q/TOF-MS/MS分析GNAS垂釣出的小分子。通過垂釣組與對照組的相對含量變化篩選26個小分子可能與GNAS有相互作用。其中7個化合物用Octet® 進一步驗證，并顯示出強的結(jié)合活性，均是SJG的主要成分。其中，黃芪中的Formononetin、高麗參的三七皂苷Ft1和人參皂苷中的甲素人參F2為“君“、地黃中的Catapol為“臣”，五味子中的五味子素A和茜草中的沒食子酸為“佐使”，符合中醫(yī)配伍“君臣佐使“原則。
 

圖5. Octet® 驗證小分子和GNAS的結(jié)合
 
非流路設計可以多次垂釣富集豐度低結(jié)合弱的小分子，提高垂釣的成功率。


中國醫(yī)學科學院基礎(chǔ)醫(yī)學研究所——脂肪肝新靶點^[4]
 
肝細胞對預防非酒精性脂肪性肝病（NAFLD）和其他慢性代謝性疾病具有重要意義。中國醫(yī)學科學院基礎(chǔ)醫(yī)學研究所黃波團隊研究發(fā)現(xiàn)，糖原合成使用其中間代謝物尿苷二磷酸葡萄糖（UDPG）來拮抗脂肪生成，從而引導小鼠和人肝細胞將葡萄糖碳儲存為糖原。在這項機制研究中，發(fā)現(xiàn)UDPG通過SLC35F5轉(zhuǎn)運進入高爾基體后，誘導site-1蛋白酶S1P的泛素化降解，從而阻斷活性形式SREBP1c的生成，最終抑制脂肪酸的合成。通過Octet® 非標記分子互作系統(tǒng)對小鼠和人的脂肪酸合成過程中關(guān)鍵酶（S1P）與UDPG結(jié)合這一重要步驟進行了驗證，并提供了結(jié)合的直觀證據(jù)。
 

圖6. 通過SA傳感器固化小鼠（a）和人（b）的S1P蛋白分別與不同濃度UDPG結(jié)合解離曲線

原文大量pulldown以及CO-IP數(shù)據(jù)，但作者仍選擇用Octet® 非標記互作系統(tǒng)來證明S1P與UDPG直接相互作用，可見Direct Binding是趨勢。
 
 
Octet® 分子互作分析系統(tǒng)的優(yōu)勢在于

• 非標記Direct Binding是趨勢，結(jié)果更準確
• 快速測定親和力，更加定量化地表征分子互作
• 無洗滌步驟，可測弱親和力（解離快）
• 寫入了美國藥典，文章多，認可度廣
• 萬金油技術(shù)，可以用與檢測DNA，小分子，蛋白質(zhì)等各種生物分子
• 多種實驗方案，除了直接測試，還有競爭法，垂釣等
• 操作簡便，耗材及維護成本低

Octet® 讓分子互作不再復雜！祝愿大家可以用賽多利斯神器發(fā)好文章！
    
《生物層干涉技術(shù)小分子應用文集》詳細介紹了生物層干涉（BLI）技術(shù)的原理，及其在小分子領(lǐng)域的多個應用場景，并對BLI技術(shù)應用于小分子研究的20余篇代表性文獻進行了整理匯總：內(nèi)容涵蓋從防御機制到垂釣驗證，再到小分子篩選、表征、優(yōu)化等多個方面，全展示了BLI技術(shù)在小分子研究中的強大潛力和實踐成果。
  
 
-參考文獻-
[1] Glucose-G protein signaling plays a crucial role in tomato resilience to high temperature and elevated CO2 | Plant Physiology | Oxford Academic (oup.com)
[2] Vaccines | Free Full-Text | Dynamic Profiling and Prediction of Antibody Response to SARS-CoV-2 Booster-Inactivated Vaccines by Microsample-Driven Biosensor and Machine Learning (mdpi.com)
[3] Zhang H, et al. Discovery of drug targets based on traditional Chinese medicine microspheres (TCM-MPs) fishing strategy combined with bio-layer interferometry (BLI) technology. Anal Chim Acta. 2024 May 29;1305:342542.
[4] Hepatic glycogenesis antagonizes lipogenesis by blocking S1P via UDPG.Science,2024

文獻列表
https://www.sciencedirect.com/science/article/abs/pii/S0376738824006367
https://www.biorxiv.org/content/10.1101/2024.06.26.600898v1.abstract
https://academic.oup.com/lifemedi/advance-article/doi/10.1093/lifemedi/lnae025/7701167
https://www.nature.com/articles/s41421-024-00699-4
https://www.tandfonline.com/doi/full/10.1080/21645515.2024.2366641
https://link.springer.com/article/10.1007/s11418-024-01825-y
https://www.sciencedirect.com/science/article/abs/pii/S0003267024007128
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.4c00242
https://pubs.acs.org/doi/abs/10.1021/acsinfecdis.3c00728
https://www.sciencedirect.com/science/article/pii/S002192582401994X
https://www.sciencedirect.com/science/article/pii/S2542529324001640
https://bpspubs.onlinelibrary.wiley.com/doi/full/10.1111/bph.16473
https://essopenarchive.org/doi/full/10.22541/au.171912146.65956164
https://journals.aai.org/jimmunol/article/doi/10.4049/jimmunol.2300706/266957
https://www.cell.com/immunity/abstract/S1074-7613(24)00279-6#%20
https://www.sciencedirect.com/science/article/pii/S2001037024002150
https://www.sciencedirect.com/science/article/abs/pii/S0168952524001331
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.3c02407
https://www.sciencedirect.com/science/article/pii/S0753332224008540
https://www.nature.com/articles/s41594-024-01319-1
https://www.sciencedirect.com/science/article/abs/pii/S1567576924010117
https://link.springer.com/article/10.1007/s40259-024-00662-5
https://www.nature.com/articles/s41467-024-49096-1
https://link.springer.com/article/10.1007/s10753-024-02060-z
https://www.science.org/doi/abs/10.1126/sciimmunol.adj5465
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.4c00555
https://onlinelibrary.wiley.com/doi/full/10.1002/advs.202401789
https://www.sciencedirect.com/science/article/abs/pii/S0965174824000821
https://www.sciencedirect.com/science/article/abs/pii/S1567576924009457
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.4c00534
https://www.sciencedirect.com/science/article/abs/pii/S0006295224003435
https://www.tandfonline.com/doi/full/10.1080/21645515.2024.2360338
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.4c00343
https://pubs.rsc.org/en/content/articlehtml/2024/sc/d4sc00267a
https://www.sciencedirect.com/science/article/abs/pii/S0043135424008091
https://www.biorxiv.org/content/10.1101/2024.06.06.597720v2.abstract
https://www.mdpi.com/2073-4468/13/2/45
https://www.cell.com/cell-reports/fulltext/S2211-1247(24)00666-1#%20
https://www.biorxiv.org/content/10.1101/2024.06.04.597356v1.full.pdf
https://www.mdpi.com/2076-2607/12/6/1148
https://www.sciencedirect.com/science/article/pii/S0753332224007108
https://www.pnas.org/doi/abs/10.1073/pnas.2400163121
https://onlinelibrary.wiley.com/doi/abs/10.1002/ange.202406677
https://www.sciencedirect.com/science/article/pii/S2590157524004176
https://link.springer.com/article/10.1007/s12020-024-03891-4
https://www.cell.com/structure/abstract/S0969-2126(24)00189-8#%20
https://www.nature.com/articles/s41420-024-02005-3
https://www.mdpi.com/1424-8247/17/6/706
https://www.sciencedirect.com/science/article/abs/pii/S0026265X24009585
https://www.cell.com/cell-reports/fulltext/S2211-1247(24)00593-X
https://www.sciencedirect.com/science/article/abs/pii/S0039914024006258
https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1387811/full
https://www.cell.com/cell-reports-medicine/fulltext/S2666-3791(24)00265-9#%20
https://journals.physiology.org/doi/epdf/10.1152/ajplung.00301.2023
https://www.nature.com/articles/s42255-024-01047-2
https://www.sciencedirect.com/science/article/pii/S0753332224006668
https://onlinelibrary.wiley.com/doi/abs/10.1002/jmv.29669
https://onlinelibrary.wiley.com/doi/abs/10.1002/jmr.3091
https://www.sciencedirect.com/science/article/abs/pii/S0308814624013888
https://www.nature.com/articles/s41467-024-48686-3
https://www.nature.com/articles/s41467-024-48504-w
https://link.springer.com/article/10.1186/s12951-024-02521-5
https://www.cell.com/cell-reports/fulltext/S2211-1247(24)00563-1#%20
https://link.springer.com/article/10.1186/s12951-024-02440-5
https://onlinelibrary.wiley.com/doi/full/10.1002/advs.202403120
https://www.sciencedirect.com/science/article/abs/pii/S0141813024031003
https://www.sciencedirect.com/science/article/abs/pii/S0141813024030423
https://www.embopress.org/doi/full/10.1038/s44318-024-00110-8
https://onlinelibrary.wiley.com/doi/full/10.1002/mco2.557
https://www.tandfonline.com/doi/full/10.1080/21505594.2024.2350893
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0302865
https://www.nature.com/articles/s41467-024-48063-0
https://www.sciencedirect.com/science/article/abs/pii/S0048357524001809
https://www.sciencedirect.com/science/article/abs/pii/S0026265X2400941X
https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1352404/full
https://www.nature.com/articles/s41467-024-47959-1
https://www.mdpi.com/1422-0067/25/9/4751
https://onlinelibrary.wiley.com/doi/abs/10.1002/adma.202402379
https://www.cell.com/cell/abstract/S0092-8674(24)00355-6#%20
https://www.biorxiv.org/content/10.1101/2024.04.20.590432v1.abstract
https://www.sciencedirect.com/science/article/abs/pii/S0378874124005105
https://www.scienceopen.com/document_file/06df5c32-226d-42ed-b05e-cc090a3d8f0f/PubMedCentral/06df5c32-226d-42ed-b05e-cc090a3d8f0f.pdf
https://www.sciencedirect.com/science/article/abs/pii/S0165993624001924
https://www.mdpi.com/2306-5354/11/4/381
https://www.cell.com/cell/abstract/S0092-8674(24)00310-6
https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1380694/full
https://www.sciencedirect.com/science/article/abs/pii/S0141813024023158
https://www.sciencedirect.com/science/article/pii/S2213231724001307
https://www.sciencedirect.com/science/article/abs/pii/S0039914024004235
https://www.nature.com/articles/s41467-024-47213-8
https://www.nature.com/articles/s41467-024-47167-x
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11128199/
https://www.nature.com/articles/s41467-024-47063-4
https://www.sciencedirect.com/science/article/pii/S0014299924002395
https://www.sciencedirect.com/science/article/abs/pii/S030881462401464X
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.4c00503
https://pubs.acs.org/doi/abs/10.1021/acs.jafc.3c04608
https://pubs.acs.org/doi/abs/10.1021/jacs.3c14046
https://www.sciencedirect.com/science/article/abs/pii/S0223523424002800
https://www.sciencedirect.com/science/article/abs/pii/S0042682224001090
https://pubs.rsc.org/en/content/articlelanding/2024/fo/d3fo03938b/unauth
https://pubs.rsc.org/en/content/articlelanding/2024/fo/d4fo00596a/unauth
https://bpspubs.onlinelibrary.wiley.com/doi/abs/10.1111/bph.16377
https://www.sciencedirect.com/science/article/abs/pii/S0168827824002770
https://www.sciencedirect.com/science/article/abs/pii/S0223523424002472
https://www.sciencedirect.com/science/article/abs/pii/S0006295224002934
https://www.sciencedirect.com/science/article/abs/pii/S1748013224001683
https://www.sciencedirect.com/science/article/abs/pii/S0142961224001984
https://link.springer.com/article/10.1007/s00262-024-03671-z
https://www.sciencedirect.com/science/article/abs/pii/S000326702400343X
https://www.sciencedirect.com/science/article/abs/pii/S1567576924004442
https://www.mdpi.com/2076-393X/12/4/352
https://www.mdpi.com/2076-0817/13/4/272
https://www.mdpi.com/1420-3049/29/7/1435
https://www.mdpi.com/2076-2607/12/3/600
https://pubs.acs.org/doi/abs/10.1021/acschembio.3c00728
https://www.mdpi.com/1420-3049/29/6/1247
https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2024.1381877/full
https://academic.oup.com/plphys/article-abstract/195/2/1025/7623298?redirectedFrom=fulltext
https://www.nature.com/articles/s41422-024-00942-3
https://www.nature.com/articles/s41467-024-46394-6
https://www.sciencedirect.com/science/article/abs/pii/S014181302401376X
https://www.cell.com/heliyon/fulltext/S2405-8440(24)04157-4#%20
https://aacrjournals.org/cancerres/article/84/10/1680/745308
https://www.sciencedirect.com/science/article/abs/pii/S014181302401465X
https://www.sciencedirect.com/science/article/abs/pii/S1386142524002841
https://www.sciencedirect.com/science/article/abs/pii/S0939641124000742
https://www.sciencedirect.com/science/article/pii/S2211383524000960
https://www.sciencedirect.com/science/article/abs/pii/S0968089624000919
https://www.sciencedirect.com/science/article/abs/pii/S0268005X24002686
https://www.sciencedirect.com/science/article/abs/pii/S0141813024018464
https://www.sciencedirect.com/science/article/abs/pii/S0141813024019317
https://www.sciencedirect.com/science/article/abs/pii/S0223523424001922
https://www.embopress.org/doi/full/10.1038/s44318-024-00076-7
https://www.tandfonline.com/doi/abs/10.2217/fvl-2023-0212
https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/abs/10.1002/bit.28707
https://academic.oup.com/ibdjournal/advance-article-abstract/doi/10.1093/ibd/izae063/7637490
https://pubs.acs.org/doi/abs/10.1021/acs.jafc.3c09598
https://pubs.acs.org/doi/abs/10.1021/acsnano.3c12783
https://link.springer.com/article/10.1186/s13071-024-06207-7
https://pubs.rsc.org/en/content/articlelanding/2024/md/d3md00752a/unauth
https://journals.asm.org/doi/abs/10.1128/aem.01851-23
https://www.cell.com/cancer-cell/fulltext/S1535-6108(24)00046-1
https://www.sciencedirect.com/science/article/abs/pii/S0223523424001508
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.3c02039
https://www.sciencedirect.com/science/article/abs/pii/S0145305X24000296
https://academic.oup.com/jac/article-abstract/79/4/903/7614994
https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/abs/10.1002/biot.202300492
https://www.sciencedirect.com/science/article/abs/pii/S0944711324000576
https://www.mdpi.com/1422-0067/25/4/2400
https://www.nature.com/articles/s41467-024-45765-3
https://www.sciencedirect.com/science/article/abs/pii/S0944711324001272
https://www.sciencedirect.com/science/article/abs/pii/S0045206824001202
https://www.sciencedirect.com/science/article/abs/pii/S1046592824000135
https://www.sciencedirect.com/science/article/abs/pii/S0223523424001077
https://www.science.org/doi/abs/10.1126/science.adi3332
https://www.sciencedirect.com/science/article/abs/pii/S0378874124002186
https://www.cell.com/molecular-therapy-family/nucleic-acids/fulltext/S2162-2531(24)00033-7#%20
https://link.springer.com/article/10.1186/s12951-024-02329-3
https://www.sciencedirect.com/science/article/pii/S2949928324000099
https://www.science.org/doi/full/10.1126/sciadv.adi9284
https://academic.oup.com/nar/article/52/5/2142/7606261
https://www.embopress.org/doi/full/10.1038/s44321-024-00026-0#tab-contributors
https://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.3c01668
https://pubs.acs.org/doi/abs/10.1021/acsbiomaterials.3c01765
https://pubs.acs.org/doi/abs/10.1021/acschembio.3c00645
https://www.sciencedirect.com/science/article/abs/pii/S0003986124000365
https://www.sciencedirect.com/science/article/abs/pii/S1748013224000380
https://journals.asm.org/doi/abs/10.1128/jvi.01515-23
https://www.sciencedirect.com/science/article/abs/pii/S0163725824000342
https://onlinelibrary.wiley.com/doi/abs/10.1002/adhm.202303619
https://www.cell.com/cell-reports/fulltext/S2211-1247(23)01664-9?uuid=uuid%3Abf3af401-1abb-4a28-ae02-e54fa34c9cdc#%20
https://www.nature.com/articles/s41401-023-01212-5
https://www.mdpi.com/1422-0067/25/13/7200
https://journals.plos.org/Plospathogens/article?id=10.1371/journal.ppat.1011948
https://www.nature.com/articles/s41467-024-45050-3
https://www.nature.com/articles/s41467-024-45202-5
https://www.nature.com/articles/s41422-023-00917-w
https://www.nature.com/articles/s41590-023-01715-7
https://www.nature.com/articles/s41467-024-44872-5#auth-Shuo-Su-Aff1
https://onlinelibrary.wiley.com/doi/full/10.1155/2024/9350822
https://onlinelibrary.wiley.com/doi/abs/10.1111/tpj.16671
https://www.cell.com/cell-reports/fulltext/S2211-1247(24)00014-7
https://journals.asm.org/doi/abs/10.1128/jvi.01954-23
https://academic.oup.com/hr/article/11/2/uhad293/7505276
https://www.tandfonline.com/doi/abs/10.4155/fmc-2023-0320
https://aacrjournals.org/cancerres/article-abstract/84/7/994/742036/Osteosarcoma-Cells-Secrete-CXCL14-That-Activates
https://www.biorxiv.org/content/10.1101/2024.01.06.574490v4.full.pdf
https://doi.org/10.1093/abt/tbad032
https://www.sciencedirect.com/science/article/pii/S2095177924000029
https://www.sciencedirect.com/science/article/pii/S2213231724000053
https://www.sciencedirect.com/science/article/abs/pii/S0168827824001065
https://www.sciencedirect.com/science/article/pii/S2090123224000407
https://www.sciencedirect.com/science/article/pii/S1756464624000331
https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2024.1348405/full
https://www.sciencedirect.com/science/article/abs/pii/S0944711324000916
https://www.sciencedirect.com/science/article/abs/pii/S1567576924000973
https://www.embopress.org/doi/full/10.1038/s44319-024-00070-4#tab-contributors
https://www.sciencedirect.com/science/article/abs/pii/S0141813024006494
https://www.sciencedirect.com/science/article/abs/pii/S0144861724001358
https://www.sciencedirect.com/science/article/abs/pii/S0168827824000424
https://pubs.acs.org/doi/abs/10.1021/acsabm.3c00936
https://pubs.acs.org/doi/abs/10.1021/acs.jafc.3c07295
https://link.springer.com/article/10.1007/s00259-023-06577-7