在每个浓度系列之前和之后注射缓冲液,并用作参考空白。
数据由Attester3.0版收集,随后在Attester Evaluation3.0版中进行处理。使用Attester Evaluation3.0版和Clamp XP获得动力学数据。对于速率常数计算,使用150kDa作为多克隆抗体的分子量。
实验结果
两种不同的多克隆抗体血清(一种针对肽,另一种针对蛋白质),及其抗原之间的相互作用所获得的数据如图1和图2所示。所用多克隆抗体的最高浓度为10和5微克/毫升,产生的频率响应分别为12和20赫兹。为了控制相互作用不受传质限制的影响,测试了几种流速,发现25升/分钟适用于相互作用研究。
在统计了所有数据,结合曲线的全局动力学分析,使用Clamp XP软件,计算出针对肽抗原的多克隆抗体的KD为7.2nM(图1),针对蛋白质抗原的多克隆抗体的KD为0.3nM(图2)。
图1:多克隆抗体与生物素化肽抗原的结合。(A)结合曲线(黑色)、结合与简单的1:1结合模型的拟合(红色)。(B)实验设置的示意图。(C)导出的速率常数和由koff /kon计算的KD 。
图2:多克隆抗体与胺偶联蛋白抗原的结合。(A)结合曲线(黑色)、结合与简单的1:1结合模型的拟合(红色)。(B)实验设置的示意图。(C)导出的速率常数和由koff /kon计算的KD 。
如上所述,多克隆抗体样品的异质性,很可能导致不同的亲和力。这也提示将简单的动力学相互作用模型与数据相匹配的可能性。图1中的A图,显示在解离阶段的第一部分有相当高的解离速度,表明亚群与抗原形成相对不稳定的复合物。此后,解离速率稳定下来,代表与抗原形成相对稳定复合物的亚群,并且从这部分解离阶段获得的数据显示与1:1相互作用模型有更好的一致性。
此外,如果使用传感器表面固定抗原的方式进行实验,为了表征二价或多价抗体,必须非常小心地测量亲和力而不要测结合总亲和力。我们以前的数据显示,使用Attana羧基或生物素传感器芯片,可以非常精确地滴定表面抗原密度,以避免由高表面密度引起的总亲和力效应。这是在上述实验开始之前对两种抗原进行的。
实验结论
1、本实验使用Attana A100 C-Fast系统,开发了一种方法,可以实时、无标记地测量多克隆抗体与多肽和蛋白质抗原的相互作用。
2、通过使用该方法,可以导出相互作用的动力学信息,例如结合(kon)和解离(koff)速率常数,并计算相互作用的亲和力。
3、该方法可用于监控多克隆抗体的纯化效率、比较不同免疫的效果、监测滴度变化等。
4、该实验可针对固定的抗原筛选多克隆抗体,节省时间且成本低。
实例4、无需纯化抗体,直接对杂交瘤上清液进行筛选和鉴定
在鉴定抗体-抗原相互作用之前纯化抗体、标记抗体,通常非常耗时,并且存在需要优化实验条件和产量低等问题。瑞典Attana蛋白互作分析仪和Attana抗体捕获试剂盒可以减少这些问题。在这里,展示了Attana的一种抗小鼠抗体的芯片,它能够从原材料中捕获抗体,随后详细研究了抗体与其抗原之间相互作用的动力学。
Attana蛋白互作分析仪,利用石英晶体微天平(QCM)技术,对分子相互作用进行实时、无标记测量。当分子被添加到传感器表面或从传感器表面移除时,共振频率的变化对应于表面质量的变化。通过将目标分子固定到传感器表面,并使相互作用的分子在表面上流动,可以实时研究相互作用。实时信息可以提供关于相互作用的动力学、亲和力和特异性数据。
实验目标
首先,从粗样品(如杂交瘤上清液)中筛选小鼠IgG抗体,确定结合抗原的解离速率。然后,确定在筛选中选择的抗原和抗体之间的详细动力学速率常数和亲和力。
实验方法
按照小鼠免疫球蛋白捕获试剂盒中提供的说明,以1xHBST(10mM HEPES缓冲盐,0.005%吐温20,25微升/分钟,22℃)为流动相,将小鼠免疫球蛋白捕获分子固定在Attana羧基传感器芯片表面,然后从杂交瘤上清液中捕获小鼠IgG抗体,然后实时研究它们各自与抗原的相互作用特性。使用5微克/毫升的抗原浓度来确定解离速率常数,使用1.25至20微克/毫升的浓度来进行详细的动力学表征。在每个相互作用循环后,表面被再生,恢复最初的小鼠IgG捕获能力。
使用Attester3.1软件收集数据,然后在Attester Evaluation3.1中进行处理。Attester Evaluation软件中嵌入的解离速率筛选工具用于识别抗原解离速率常数最低的抗体。使用Attester Evaluation和Clamp XP软件计算和获得详细的动力学数据。
实验结果
针对一种抗原,从产生小鼠IgG的杂交瘤培养物中筛选了11种抗体的解离速率常数。首先,使用小鼠IgG结合分子从各自的上清液中捕获IgG。此后,将抗原注射到表面上,并监测与捕获的抗体的相互作用。使用Attester Evaluation软件的解离速率筛选工具用于生成一个列表,突出显示最稳定的相互作用(低解离速率常数)。参见图1的结果图和收集的数据。
图1、11种不同抗体的抗原相互作用曲线汇总。对于每个相互作用周期,首先注射一个含有特定抗体的杂交瘤上清液以捕获抗体,随后注射抗原。图中还显示了Attester Evaluation软件的解离速率筛选工具。该表给出了11个确定的解离常数。阈值功能可以用于突出显示解离速率常数低于某个值的抗体。如图所示,低于1.8 x 10-4的解离速率常数以蓝色突出显示 。
选择具有最低解离速率的杂交瘤,用于进一步抗体-抗原相互作用的详细动力学分析。在该步骤中,使用了与解离速率筛选相同的芯片。在保持抗体捕获水平恒定的同时,改变抗原浓度,通过使用全局曲线拟合算法分析数据来得到详细的动力学速率常数(即相互作用的kon、koff和KD,图2)
图2、(A)动力学速率常数kon 和koff的测定。相互作用数据用黑色表示,动力学模型的拟合用红色表示。(B)捕获分析实验设置的示意图。(C)速率常数。
实验结论
1、利用Attana蛋白互作分析仪,开发了一种实时、无标记的从杂交瘤上清液中捕获小鼠抗体的方法。同样的方法可以用于捕获人类免疫球蛋白。
2、导出相互作用、结合(Kon)和解离(Koff)速率常数的动力学信息,可以计算分子之间的亲和力(KD)。
3、实现了直接从粗样品(如杂交瘤上清液)中快速筛选多达192个抗体-抗原对,而无需纯化或标记抗体。
4、该方法使用通用的固定和再生条件,并且不需要预先纯化抗体,可以显著节省时间、提高效率。
应用Attana Cell 200蛋白互作分析仪的参考文献
1.The mechanism of Hsp90-induced oligomerizaton of Tau. Weickert S, Wawrzyniuk M, John LH, Rüdiger SGD, Drescher M. Science Advances, Mar 13;6(11).2020
2.The TLR9 Agonist Cobitolimod Induces IL10-Producing Wound Healing Macrophages and Regulatory T Cells in Ulcerative Colitis. Heike Schmitt, Julia Ulmschneider, Ulrike Billmeier, Michael Vieth, Patrizio Scarozza, Sophia Sonnewald, Stephen Reid, Imke Atreya, Timo Rath, Sebastian Zundler, Melanie Langheinrich, Jürgen Schüttler, Arndt Hartmann, Thomas Winkler, Charlotte Admyre, Thomas Knittel, Christine Dieterich Johansson, Arezou Zargari, Markus F Neurath, Raja Atreya. Journal of Crohn's and Colitis, 1-17, 2019
3.Recombinant spider silk coatings functionalized with enzymes targeting bacteria and biofilms. Fredrik Seijsing. Linnea Nilebäck. Oskar Öhman. Rajeev Pasupuleti. Camilla Ståhl, Johan Seijsing, My Hedhammar. Microbiology Open, December 2019
4.Electro-Responsive Suce Composition and Kinetics of an Ionic Liquid in a Polar Oil. Nicklas Hjalmarsson, Erik Bergendal, Yong-Lei Wang, Bulat Munavirov, Daniel Wallinder, Sergei Glavatskih, Teodor Aastrup, Rob Atkin, István Furó, Mark W. Rutland. Langmuir, 2019, 35, 48, 15692-15700
5.Monovalent TNF receptor 1-selective antibody with improved affinity and neutralizing activity. Fabian Richter, Oliver Seifert, Andreas Herrmann, Klaus Pfizenmaier, Roland E. Kontermann. mAbs, 11 (1), 2019, p. 166-177
6.Dectin-1 Binding to Annexins on Apoptotic Cells Induces Peripheral Immune Tolerance via NADPH Oxidase-2. Kevin Bode et al.. Cell Rep, Dec 24;29(13):4435-4446.2019
7.Phenotypic Screen with the Human Secretome Identifies FGF16 as Inducing Proliferation of iPSC-Derived Cardiac Progenitor Cells. Karin Jennbacken, Fredrik Wågberg, Ulla Karlsson et al.. International Journal of Molecular Sciences 06.12.19
8.Rapid affinity chromatographic isolation method for LDL in human plasma by immobilized chondroitin-6-sulfate and anti-apoB-100 antibody monolithic disks in tandem. Thana Liangsupree, Evgen Multia, Jari Metso, Matti Jauhiainen, Patrik Forssén, Torgny Fornstedt, Katariina Öörni, Aleš Podgornik & Marja-Liisa Riekkola. Scientific Reportsvolume 9, Article number: 11235 (2019)
9.Defining the structural basis for human alloantibody binding to human leukocyte antigen allele HLA-A*11:01. Yue Gu, Yee Hwa Wong, Chong Wai Liew et al.. Nature Communications, 10 (893), 2019
10.Improved monovalent TNF receptor 1-selective inhibitor with novel heterodimerizing Fc. Fabian Richter, Oliver Seifert, Andreas Herrmann, Klaus Pfizenmaier, Roland E. Kontermann. mAbs, 31 of March 2019
11.Anti-LRP5/6 VHHs promote differentiation of Wnt-hypersensitive intestinal stem cells. Nicola Fenderito, David Proverbio et al.. Nature Communications, 10 (365).2019
12.Monovalent TNF receptor 1-selective antibody with improved affinity and neutralizing activity. Fabian Richter, Oliver Seifert, Andreas Herrmann, Klaus Pfizenmaier, Roland E. Kontermann. mAbs, 11 (1), 2019, p. 166-177
13.Generation by phage display and characterization of drug-target complex-specific antibodies for pharmacokinetic analysis of biotherapeutics. Harth S, Ten Haaf A, Loew C, Frisch C, Knappik A. MAbs. 2019 Jan;11(1):178-190.
14.A Single-Chain-Based Hexavalent CD27 Agonist Enhances T Cell Activation and Induces Anti-Tumor Immunity. Thiemann M et al.. Front Oncol. 2018 Sep 19;8:387.
15.Fabrication of Carbohydrate Chips Based on Polydopamine for Real-Time Determination of Carbohydrate–Lectin Interactions by QCM Biosensor. Kun Sang et al.. Polymers 2018, 10(11), 1275
16.Differential recognition of Haemophilus influenzae whole bacterial cells and isolated lipooligosaccharides by galactosespecific lectins. Ioanna Kalograiaki, Begoña Euba, María del Carmen et al.. Nature, Scientific Reports 8, 16292 (2018)
17.Selective Sensing of the Biotinyl Moiety Using Molecularly Imprinted Polyaniline Nanowires. S. Mandal, S. Suriyanarayanan, I. A. Nicholls & K.Ramanujam. J. Electrochem. Soc. 2018 volume 165, issue 14,
18.Effects of the flow type on the immobilization of horseradish peroxidase on polymers. Spaska Yaneva, Tzvetan Velinov, Lyubov Yotova.. Journal of Chemical Technology and Metallurgy, vol. 53 (5), 2018, p. 791-800.
19.Label-Free Cell-Based Assay for Characterization of Biomolecules and Receptors. Diluka Peiris, Lars M. Jørgensen et al.. Epitopte Mapping Protocols, 2018, p. 53-63
20.Bacterial Suce Glycans: Microarray and QCM Strategies for Glycophenotyping and Exploration of Recognition by Host Receptors.. I. Kalograiaki, M Campanero-Rhodes, D. Proverbio, B. Euba, J. Garmendia, T. Aastrup et al.. Methods in Enzymology, vol. 598, 2018, p. 37-70
21.Off-target testing assays. Samuel Altun & Teodor Aastrup.. Scientistlive, 23rd January 2018.
22.Reliable Strategy for Analysis of Complex Biosensor Data. P. Forssén, J. Samuelsson, T. Aastrup, S. Altun, D. Wallinder, L. Wallbing, T. Fornstedt et al.. Analytical Chemistry, vol. 90 (8), 2018, p. 5366-5374.
23.Label-free in-flow detection of receptor recognition motifs on the biomolecular corona of nanoparticles. M. Gianneli, E. Polo, H. Lopez, V. Castagnola, T. Aastrup & K.A. Dawson.. Nanoscale, vol. 10, 2018, p. 5474-5481.
24.Virus-like particle display of HER2 induces potent anti-cancer responses. Arianna Palladini, Susan Thrane, Jessica Pihl, Adam F. Sander et al.. OncoImmunology, vol 7 (3), 2018,
25.Electrochemically synthesized molecularly imprinted polythiophene nanostructures as recognition elements for an aspirin-chemosensor. Subramanian Suriyanarayanana, Ian A. Nicholls et al.. Sensors and Actuators B: Chemical. Volume 253, December 2017, Pages 428-436
26.Synthesis and binding affinity analysis (..) using quartz crystal microbalance sensors. Oscar Norberg, Bin Wu, Olof Ramström et al.. Carbohydrate Research, vol. 452, 2017, p. 35-42.
27.QCM Biosensor Based on Polydopamine Suce for Real-Time Analysis of the Binding Kinetics of Protein-Protein Interactions. Chunli Wu, Xueming Li,Siyu Song et al.. Polymers, vol. 9(10), 2017, p. 482.
28.Optimization of 3D Suces of Dextran with Different Molecule Weights for Real-Time Detection of Biomolecular Interactions by a QCM Biosensor. Siyu Song, Yuchao Lu, Xueming Li et al.. Polymers, vol. 10(10), 2017, p. 409.
29.Novel QCM based method to predict the in-vivo behaviour of nanoparticles. M. Gianneli Y. Yan, E. Polo, D. Peiris et al.. Procedia Technology, vol. 27, 2017, p. 197-200.
30.Novel strategies to mimic transmembrane tumour necrosis factor-dependent activation of tumour necrosis factor receptor 2. Roman Fischer, Jessica Marsal, Cristiano Guttà, Roland E. Kontermann et al.. Scientific Reports, vol. 7, article 6607, 2017.
31.Inhibition of HER3 activation and tumour growth with a human antibody binding to a consered epitope formed by domain III and IV. Lisa C. Schmitt, Alexander Rau, Oliver Seifert, Jonas Honer, Roland E. Kontermann et al.. mAbs, vol. 9, 2017, p. 831-843.
32.Temperature effect on the complex formation between Plluronic acid F127 and starch. Yana Petkova-Olsson, Samuel Altun, Henrik Ullsten, Lars Janstrom. Carbohydrate Polymers, vol. 166, p. 264-270, 2017.
33.Cellular glycosylation affects Herceptin binding and sensitivity of breast cancer cells to doxorubicin and growth factors. Diluka Peiris, Alexander F. Spector, Hazel Welch, Miriam V. Dwek et al.. Scientific Reports, vol. 7, article 43006, 2017.
34.Selection of a monoclonal antibody by ELISA, immunoblotting and Quartz Crystal Microbalance technology for immunohistochemical detection of Mycoplasma mycoides subsp. mycoide. Luciani M, Armillotta G, Manna L, Ciarelli A, Di Febo T, Krasteva I, Pini A, Sacchini F, D'Angelo AR. Vet Ital. 2016 Jul 15.
35.Fluorescent Image–Guided Surgery with an Anti-Prostate Stem Cell Antigen (PSCA) Diabody Enables Targeted Resection of Mouse Prostate Cancer Xenografts in Real Time. Geoffrey A. Sonn, Andrew S. Behesnilian, Ziyue Karen Jiang, et al.. Clinical Cancer Research, vol. 22 (6), 2016, p.
36.Spatially Well-Defined Carbohydrate Nanoplatforms: Synthesis, Characterization and Lectin Interaction Study. B.J.J. Timmer D. Proverbio, S. Altun, O. Ramström, T. Aastrup, S.P. Vincent et al.. Chem. Commun., vol. 52, p. 12326-12329, 2016.
37.Signal enhancement in ligand-receptor interactions using dynamicpolymers at quartz crystal microblance sensors. Gunnar Dunér, Henrik Anderson, Zhichao Pei, Bjorn Ingemarsson et al.. Analyst, vol. 141, p. 3993-3996, 2016.
38.Combined Bacteria Microarray and Quartz Crystal Microbalance Approach for Exploring Glycosignatures…. Ioanna Kalograiaki, Begoña Euba, Davide Proverbio et al.. Anal. Chem, vol. 88 (11), p. 5950–5957, 2016.
39.Real-time and label free determination of ligand binding-kinetics to primary cancer tissue specimens…. Thomas Mandel Clausena, Marina Ayres Pereiraa et al.. Sensing & Bio-Sensing Research vol. 9, p. 23-30, 2016.
40.Glyconanomaterials for biosensing applications. Nanjing Hao, Kitjanit Neranon, Olof Ramström, Mingdi Yan. Biosensors and Bioelectronics, vol. 76, p. 113-130, 2016.
41.Facile fabrication of glycopolymer-based iron oxide nanoparticles and their applications…. Chen Shao, Xumig Le, Zhichao Pei, Li,a Zhichao Pei et al.. Polym. Chem., vol. 7, p. 1337-1344, 2016.
42.Ionization of covalent immobilized poly(4-vinylphenol) monolayers measured by ellipsometry, QCM and SPR. Suji Uppalapati, Na Kong, Oscar Norberg, Olof Ramström,∗, Mingdi Yan. Applied Suce Science, vol. 343, p. 166-171, 2015.
43.Pharmacokinetic properties of IgG and various Fc fusion proteins in mice. Felix Unverdorben, Fabian Richter, Meike Hutt, Oliver Seifert et al.. mAbs , vol. 8(1), p. 120-128, 2016.
44.Oriented and reversible immobilization of His-tagged proteins on two- and three-dimensional suces…. Xueming Lia, Siyu Songa, Yuxin Peia, Hai Dongb, , Teodor Aastrupc, Zhichao Peia.. Sensors and Actuators B: Chemical, vol. 224, p. 814-822, 2016.
45.Biomimetic Approach for Ion Channels Based on Suctant Encapsulated Spherical Porous Metal-Oxide Capsules. Eugene Mahon, Somenath Garai, Achim Müller & Mihail Barboiu.. Adv Mater, vol. 27(35), p. 5165-5170, 2015.
46.Targeting Human Cancer by a Glycosaminoglycan binding Malaria protein. Ali Salanti, Thomas M. Clausen, Mette Ø. Agerbæk et al.. Cancer Cell, vol. 28 (4), p. 500–514, 2015.
47.A Fab-Selective Immunoglobulin-Binding Domain from Streptococcal Protein G with Improved Half-Life Extension Properties. Felix Unverdorben, Meike Hutt, Oliver Seifert, Roland E. Kontermann. PLoS ONE, vol. 10(10), 2015.
48.Weighing the suce charge of an ionic liquid. Nicklas Hjalmarsson, Daniel Wallinder, Sergei Glavatskih, Rob Atkin et al.. Nanoscale, vol. 7, p. 16039-16045, 2015.
49.Real time and label free analysis of binding thermodynamics of carbohydrate protein interactions…. Xueming Li1,Siyu Song, Qi Shuai,Yihan Pei, Teodor Aastrup, Yuxin Pei & Zhichao Pei. Scientific Reports 5, article 14066, 2015.
50.Synthesis and binding affinity analysis of positional thiol analogs of mannopyranose for the elucidation of sulfer in different position. Bin Wu, Jiantao Ge, Bo Ren, Zhichao Pei, Hai Dong. Tetrahedron, vol. 71 (23), p. 4023-4030, 2015.
51.Investigating manufactured nanoparticles. Teodor Aastrup et al.. Biotechnology, June 2015
52.Study of the Interaction of Trastuzumab and SKOV3 epithelial cancer cells using a Quartz Crystal Microbalance Sensor. Louise Elmlund, Camilla Käck, Teodor Aastrup & Ian A. Nicholls. Sensors, vol. 15(3), p. 5884-5894, 2015.
53.Preclinical and early clinical development of GNbAC1, a humanized IgG4 monoclonal antibody targeting endogenous retroviral MSRV-Env protein.. François Curtin, Hervé Perron, Arno Kromminga, Hervé Porchet & Alois B Lang. mAbs , vol. 7(1), p. 265-275, 2015.
54.One-Step Synthesis of Dual Clickable Nanospheres via Ultrasonic…. Yong Hou, Shoupeng Cao, Xueming Li, Beibei Wang, Yuxin Pe, Lin Wang, & Zhichao Pei. ACS Appl. Mater. Inteces, 2014, 6 (19), pp 16909–16917
55.3D modeling and characterization of the human CD115 monoclonal antibody H27K15 epitope and design of a chimeric CD115 target. Benoît Grellier, Fabrice Le Pogam, Marc Vitorino et al.. MAbs. 2014 Mar-Apr;6(2):533-46
56.Improved Modeling of In Vivo Kinetics of Slowly Diffusing Radiotracers for Tumor Imaging. Moses Q. Wilks, Scott M. Knowles, Anna M. Wu and Sung-Cheng Huang. J Nucl Med, vol. 55(9), p. 1539-44, 2014.
57.Quantitative ImmunoPET of Prostate Cancer Xenografts with 89Zr- and 124I-Labeled Anti-PSCA A11 Minibody. Scott M. Knowles, Kirstin A. Zettlitz, Richard Tavaré et al.. J Nucl Med, vol. 55(3), p. 452–459, 2014.
58.Highly potent dUTPase inhibition by a bacterial repressor protein reveals a novel mechanism for gene expression control. Judit E. Szabó, Veronika Németh, Veronika Papp-Kádár et al.. Nucleic Acids Res. 2014 Oct 29; 42(19): 11912–11920.
59.Soluble components of the flagellar export apparatus, FliI, FliJ, and FliH, do not deliver flagellin (…) from the cytosol to the export gate. Ráchel Sajó, , Károly Liliom , , Adél Muskotál , Ágnes Klein et al.. Molecular Cell Research, vol. 1843 (11) p. 2414-2423, 2014
60.Mapping the immunogenicity of therapeutic protein products. Samuel Altun, Lena Heffler and Teodor Aastrup. Scientistlive, 30th May 2014.
61.Oligosaccharide G19 inhibits U-87 MG human glioma cells growth in vitro and in vivo by targeting epidermal growth factor (EGF)…. Hailing Liu Ling Zhou, Songshan Shi, Ying Wang,Xinyan Ni et al.. Glycobiology, vol. 24 (8), p. 748–765, 2014.
62.Label free cell based assay for the characterisation of peptide receptor interaction. Davide Proverbio, Jana Valnohova, Shane Wright, Gunnar Schulte and Teodor Aastrup. Clinical Research, vol. 6(2), 2014.
63.Partial-filling affinity capillary electrophoresis and quartz crystal microbalance with adsorption energy distribution calculations…. Lipponen, Tähkä, Samuelsson,Jauhiainen, Metso, Cilpa-Karhu, Fornstedt, Kostiainen, Riekkola. Anal Bioanal Chem. 2014 Jul;406(17):4137-46
64.Hierarchical Thin Film Architectures for Enhanced Sensor Performance: Liquid Crystal-Mediated Electrochemical Synthesis…. Subramanian Suriyanarayanan, Hazrat Nawaz, Natacha Ndizeye, and Ian A. Nicholls. Biosensors 2014, 4(2), 90-110
65.A Phage Display Screening Derived Peptide with Affinity for the Adeninyl Moiety. Elmlund, Söderberg, Suriyanarayanan, Nicholls. Biosensors 2014, 4(2), 137-149
66.Entropy-driven lectin-recognition of multivalent glycovesicles. Zineb Mouline, Eugene Mahon, Emeline Gomez, Veronique Barragan-Montero et al.. Chemical Communications, Issue 6, 2014.
67.Atmospheric pressure plasma polymers fortuned QCM detection of protein adhesion. Rusu, Asandulesa, Topala, Pohoata, Dumitrascu, Barboiu. Biosens Bioelectron. 2014 Mar 15;53:154-9
68.Biotin selective polymer nano-films. Elmlund, Suriyanarayanan, Wiklander, Aastrup, Nicholls. J Nanobiotechnology. 2014 Mar 21;12:8
69.Multivalent Recognition of Concanavalin A by {Mo132} Glyconanocapsules-Toward Biomimetic Hybrid Multilayers. Barboiu, Mouline, Silion, Licsandru, Simionescu, Mahon, Pinteala. Chemistry - A European JournalVolume 20, Issue 22, 2014.
70.Inducement of Cytokine Release by GFPBW2, a Novel Polysaccharide from Fruit Bodies of Grifola frondosa, through Dectin‑1 in Macrophages. Ying Wang, Jianping Fang, Xinyan Ni, Jie Li, Qin Liu, Qun Dong et al.. J Agric Food Chem, vol. 61(47), p. 11400-11409, 2013.
71.Binding to Heparan Sulfate Chains Modulates Cellular Activity by Selectively The C-terminal Peptide of Chondroadherin. Haglund L, Tillgren V, Önnerfjord P & Heinegård D.. J Biol Chem, vol. 288(2), p. 995-1008, 2013.
73.Three complementary techniques for the clarification of temperature effect on low-density lipoprotein–chondroitin-6-sulfate interaction. Cilpa-Karhu, Lipponen, Samuelsson, Öörni, Fornstedt, Riekkola. Anal Biochem. 2013 Dec 15;443(2):139-47
74.A unique anti-CD 115 monoclonal antibody which inhibits osteolysis and skews human monocyte differentiation…. Haegel H, Thioudellet C, Hallet R, Geist M, Menguy T, Le Pogam F et al.. MAbs. 2013 Sep-Oct;5(5):736-47
75.Antagonistic TNF Receptor One-Specific Antibody (ATROSAB): Receptor Binding and In Vitro Bioactivity. Richter, Liebig, Guenzi, Herrmann, Scheurich, Pfizenmaier, Kontermann. Plos One, August 19, 2013.
76.An anti-TNFR1 scFv-HSA fusion protein as selective antagonist of TNF action. Verena Berger, Fabian Richter, Kirstin Zettlitz, Felix Unverdorben et al.. Protein Eng Des Sel. 2013 Oct;26(10):581-7
77.Protection by immunoglobulin dual-affinity retargeting antibodies against dengue virus. Brien, Sukupolvi-Petty, Williams, Lam, Schmid, Johnson, Harris, Diamond. J Virol. 2013 Jul;87(13):7747-53
78.Protein-resistant hyperbranched polyethyleneimine brush suces. Subramanian Suriyanarayanan, Hung-Hsun Lee, Bo Liedberg, Teodor Aastrup, Ian A. Nicholls. J Colloid Intece Sci. 2013 Apr 15;396:307-15
79.A suspension-cell biosensor for real-time determination of binding kinetics of protein–carbohydrate interactions on cancer cell suces. Xueming Li, Yuxin Pei, Ruina Zhang, Qi Shuai, Feng Wang, Teodor Aastrup and Zhichao Pei. Chemical Communications, Issue 85, 2013.
80.Photogenerated lectin sensors produced by thiol-ene/yne photo-click chemistry in aqueous solution. Oscar Norberg, Irene H. Lee, Teodor Aastrup, Mingdi Yan and Olof Ramström. Biosens Bioelectron. 2012 Apr 15;34(1):51-6
81.A novel approach to determining the affinity of protein–carbohydrate interactions employing adherent cancer cells grown on a biosensor suce. Diluka Peiris, Anatoliy Markiv, G. Paul Curley, Miriam V. Dwek. Biosens Bioelectron. 2012 May 15;35(1):160-6
82.Functionalization of single-walled carbon nanotubes and their binding to cancer cells. S.Y. Madani, A. Tan, M. Dwek, A. Seifalian. Int J Nanomedicine. 2012;7:905-14
83.A New Generation of Cell-Biosensors. Miriam Dwek and Teodor Aastrup. The Pharmaceutical Technology Journal.2012
84.Real-time analysis of the carbohydrates on cell suces using a QCM biosensor: a lectin-based approach. Z- Pei, J. Saint-Guirons, C. Käck, B. Ingemarsson, T. Aastrup. Biosens Bioelectron. 2012 May 15;35(1):200-5
85.Domain-based assays of individual antibody concentrations in an oligoclonal combination targeting a single protein. Q. Meng, M. Li, M.A. Silberg, F. Conrad, J. Bettencourt et al.. Analytical Biochemistry, vol. 421(2), p. 351-361, 2012.
86.Half-life extension of a single-chain diabody by fusion to domain B of staphylococcal protein A. Unverdorben, Färber-Schwarz, Richter, Hutt, Kontermann. Protein Engineering, vol. 25(2), p. 81–88, 2012.
87.The effect of adjuvants on the immune response induced by a DBL4ɛ-ID4 VAR2CSA based Plasmodium falciparum vaccine against placental malari. Pinto VV, Salanti A, Joergensen LM, Dahlbäck M, Resende M et al.. Vaccine Volume 30, Issue 3, 11 January 2012, Pages 572-579
88.Structural and Functional Insight into How the Plasmodium falciparum VAR2CSA Protein Mediates Binding…. Thomas M. Clausen, Stig Christoffersen Madeleine Dahlbäck, Annette Eva Langkilde. J Biol Chem. 2012 Jul 6; 287(28):23332-45
89.Plasma half-life extension of small recombinant antibodies by fusion to immunoglobulin-binding domains (IgBD). Meike Hutt, Aline Färber-Schwarz, Felix Unverdorben et al.. J Biol Chem. 2012 Feb 10; 287(7): 4462–4469.
90.Heparin Impairs Angiogenesis through Inhibition of MicroRNA-10b. Xiaokun Shen, Jianping Fang, Xiaofen Lv, Zhicao Pei et al.. J Biol Chem, vol 286(30), p. 26616–26627, 2011.
91.In Vitro Evolution of Allergy Vaccine Candidates, with Maintained Structure, but Reduced B Cell and T Cell Activation Capacity. Ola B. Nilsson, Justus Adedoyin, Claudio Rhyner, Theresa Neimert-Andersson et al.. Plos one, September 13, 2011.
92.Unbinding of hyaluronan accelerates the enzymatic activity of bee hyaluronidase. Iliás, Liliom, Greiderer-Kleinlercher, Reitinger, Lepperdinger. J Biol Chem. 2011 Oct 14; 286(41): 35699–35707.
93.Neonatal FcR Overexpression Boosts Humoral Immune Response in Transgenic Mice. Cervenak, Bender, Schneider, Magna, Valer Carstea, Liliom, Erdei, Bosze and Kacskoics. The Journal of ImmunologyVol. 186, Issue 2, 15 Jan 2011
94.Polyethylene glycol-stabilized lipid disks as model membranes in interaction studies based on electro kinetic capillary chromatography…. K. Vainikka, K. Reijmar, G. Yohannes, J. Samuelsson, K. Edwards et al.. Anal Biochem. 2011 Jul 1;414(1):117-24
95.Collagen I and III and their decorin modified suces studied by atomic force microscopy and the elucidation of their affinity toward.... Witos J, Saint-Guirons J, Meinander K, D'Ulivo L, Riekkola ML. Analyst. 2011 Sep 21;136(18):3777-82
96.Improving the Developability of Biopharmaceuticals. Zurdo J, Michael R, Stallwood Y, Hedman K and Aastrup T. Discovery Technology, 2011.
97.Magnetosome expression of functional camelid antibody fragments (nanobodies) in Magnetospirillum gryphiswaldense. Pollithy, Romer, Lang, Müller, Helma, Leonhardt, Rothbauer, Schüler. Appl Environ Microbiol. 2011 Sep; 77(17): 6165–6171.
98.Towards a Synthetic Avidin Mimic. Wiklander J, Karlsson BCG, Aastrup T, Nicholls IA. Anal. & Bioanal. Chemistry, vol. 400(5), p. 1397–1404, 2011.
99.Three Different Approaches for the Clarification of the Interactions between Lipoproteins and Chondroitin-6-sulfate. K. Lipponen, P.W. Stege, G. Cilpa, J. Samuelsson, T. Fornstedt and M.-L. Riekkola. Anal. Chem., 2011, 83 (15), pp 6040–6046