Tailored molecular interaction analysis, data processing and help with professional reporting
What service do you offer?
At the Molecular Interactions Core Facility, we provide a full service, meaning that you deliver the interaction partners and leave the rest to us:
- Tailor-made assays. With a little help from your expert knowledge about the molecules in question, we design an assay optimized to deliver the kind of data you desire.
- Sample analysis. We optimize the experimental conditions and run the samples.
- Data processing and interpretation. We process your data, and will happily discuss how they should be interpreted.
- Reporting. If you wish, we will help you making good-looking figures and writing M&M sections for publication.
Who can use this service?
The Molecular Interactions Core Facility at the Department of Biomedicine offers molecular interaction analysis services for all scientists at Aarhus University as well as external academic and industrial partners.
Equipment
Biacore 3000
SPR analysis, which depends on surface immobilization of one of the interacting molecules, is performed on a Biacore 3000 instrument. In addition to affinity constants (KD), this technique also allows determination of kinetic rate constants ka and kd (association and dissociation rates).
Applications
Surface Plasmon Resonance, on the Biacore platform, is most frequently used for binding/kinetic analysis of interactions between protein-protein or protein-small molecule. However, it can detect binding between any kinds of molecules, provided that one of them (termed the ligand in SPR terminology) can be immobilized on the chip surface either covalently or non-covalently. As listed below, SPR has also been employed for a number of other applications.
- Binding analysis - binding or no binding
- Kinetic analysis - determination of affinity (KD) and rate constants ka and kd
- Concentration measurements - determination of analyte concentration
- Identification of interaction partners - bound analyte protein can be recovered and identified by mass spectrometry
- Epitope mapping
Highlights
- Low sample consumption. Often a few micrograms of both analyte and ligand is sufficient for a full kinetics experiment.
- Kinetic parameters ka and kd can be obtained.
- A broad range of affinities (mM to pM).
- Ligand needs to be immobilized, either covalent or non-covalently.
Price
Customers will be charged by the hour the instrument is running and of course for the consumables provided by us.
At the facility, we always keep stocks of the most common standard Biacore consumables such as CM5 sensor chips.
| Prices per hour (or part thereof) | |||
| Academic customers | 250 | ||
| Private companies | 550 | ||
Standard consumables | |||
| 1,200 | ||
| 500 | ||
| 1,800 | ||
| 500 | ||
*Sensor chips for antibody capture can be “shared” by many users, studying diverse molecular interactions. These sensor chip surfaces can usually be regenerated at least 100 times, allowing for several experiments to be performed on the same chip.
Price example 1
The customer requested a kinetics experiment for a previously uncharacterized protein-protein interaction. The experiment was carried out on a “shared” protein G antibody capture chip, greatly reducing the time spend on assay optimization. Setting up and optimizing the assay took 4 hours. Capture antibody, ligand and analyte were provided by the customer. Eight different concentrations of analyte were analyzed + appropriate number of blank runs:
| Consumables, protein G sensor chip (shared) | 500 |
| Experimental setup optimization, 4 hours | 1000 |
| Kinetics run, 5 hours | 1,250 |
| Total | 2,750 |
Price example 2
The customer requested a kinetics experiment for a previously uncharacterized protein-protein interaction. An appropriate level of ligand was immobilized on a new CM5 chip. Due to the unknown nature of this interaction, considerable time was spend optimizing the regeneration conditions. Hence, ligand immobilization and assay optimization took a total of eight hours. Ligand and analyte were provided by the customer. Eight different concentrations of analyte were analyzed + appropriate number of blank runs. The kinetics run itself took only four hours, as this setup requires fewer steps per cycle:
| Consumables, CM5 sensor chip | 1,200 |
| Ligand immobilization, 2 hours | 500 |
| Experimental setup optimization, 6 hours | 1,500 |
| Kinetics run, 4 hours | 1,000 |
| Total | 4,200 |
Booking
Write an email to Biacore.biomed@au.dk
Location
Biacore – Molecular Interactions Core Facility
Department of Biomedicine
Bld 1234, 217A
Wilhelm Meyers Allé 3
8000 Aarhus C
Denmark
Publications and acknowledgements
Acknowledgements
Please acknowledge the Biacore – Molecular Interactions Core Facility, when publishing results obtained using our services, and please advise us about the publication by e-mailing us a link to the publication at biacore.biomed@au.dk.
Publications
Birn H, Verroust PJ, Nexo E, Hager H, Jacobsen C, Christensen EI and Moestrup SK. Characterization of an epithelial approximately 460-kDa protein that facilitates endocytosis of intrinsic factor-vitamin B12 and binds receptor-associated protein. J Biol Chem 272: 26497-26504, 1997.
Graversen JH, Lorentsen RH, Jacobsen C, Moestrup SK, Sigurskjold BW, Thogersen HC and Etzerodt M. The plasminogen binding site of the C-type lectin tetranectin is located in the carbohydrate recognition domain, and binding is sensitive to both calcium and lysine. J Biol Chem 273: 29241-29246, 1998.
Hartmann R, Norby PL, Martensen PM, Jorgensen P, James MC, Jacobsen C, Moestrup SK, Clemens MJ and Justesen J. Activation of 2'-5' oligoadenylate synthetase by single-stranded and double-stranded RNA aptamers. J Biol Chem 273: 3236-3246, 1998.
Kozyraki R, Kristiansen M, Silahtaroglu A, Hansen C, Jacobsen C, Tommerup N, Verroust PJ and Moestrup SK. The human intrinsic factor-vitamin B12 receptor, cubilin: molecular characterization and chromosomal mapping of the gene to 10p within the autosomal recessive megaloblastic anemia (MGA1) region [published erratum appears in Blood 1998 Oct 1;92(7):2608]. Blood 91: 3593-3600, 1998.
Moestrup SK, Schousboe I, Jacobsen C, Leheste JR, Christensen EI and Willnow TE. beta2-glycoprotein-I (apolipoprotein H) and beta2-glycoprotein-I- phospholipid complex harbor a recognition site for the endocytic receptor megalin. J Clin Invest 102: 902-909, 1998.
Tauris J, Ellgaard L, Jacobsen C, Nielsen MS, Madsen P, Thogersen HC, Gliemann J, Petersen CM and Moestrup SK. The carboxy-terminal domain of the receptor-associated protein binds to the Vps10p domain of sortilin. FEBS Lett 429: 27-30, 1998.
Christensen EI, Moskaug JO, Vorum H, Jacobsen C, Gundersen TE, Nykjaer A, Blomhoff R, Willnow TE and Moestrup SK. Evidence for an essential role of megalin in transepithelial transport of retinol. J Am Soc Nephrol 10: 685-695, 1999.
Hilpert J, Nykjaer A, Jacobsen C, Wallukat G, Nielsen R, Moestrup SK, Haller H, Luft FC, Christensen EI and Willnow TE. Megalin antagonizes activation of the parathyroid hormone receptor. J Biol Chem 274: 5620-5625, 1999.
Kozyraki R, Fyfe J, Kristiansen M, Gerdes C, Jacobsen C, Cui S, Christensen EI, Aminoff M, de lC, Krahe R, Verroust PJ and Moestrup SK. The intrinsic factor-vitamin B12 receptor, cubilin, is a high-affinity apolipoprotein A-I receptor facilitating endocytosis of high-density lipoprotein. Nat Med 5: 656-661, 1999.
Kristiansen M, Kozyraki R, Jacobsen C, Nexo E, Verroust PJ and Moestrup SK. Molecular dissection of the intrinsic factor-vitamin B12 receptor, cubilin, discloses regions important for membrane association and ligand binding [In Process Citation]. J Biol Chem 274: 20540-20544, 1999.
Leheste JR, Rolinski B, Vorum H, Hilpert J, Nykjaer A, Jacobsen C, Aucouturier P, Moskaug J, Otto A, Christensen EI and Willnow TE. Megalin Knockout Mice as an Animal Model of Low Molecular Weight Proteinuria. Am J Pathol 155: 1361-1370, 1999.
Munck PC, Nielsen MS, Jacobsen C, Tauris J, Jacobsen L, Gliemann J, Moestrup SK and Madsen P. Propeptide cleavage conditions sortilin/neurotensin receptor-3 for ligand binding. EMBO J 18: 595-604, 1999.
Nielsen MS, Jacobsen C, Olivecrona G, Gliemann J and Petersen CM. Sortilin/neurotensin receptor-3 binds and mediates degradation of lipoprotein lipase. J Biol Chem 274: 8832-8836, 1999.
Niemeier A, Willnow T, Dieplinger H, Jacobsen C, Meyer N, Hilpert J and Beisiegel U. Identification of megalin/gp330 as a receptor for lipoprotein(a) in vitro. Arterioscler Thromb Vasc Biol 19: 552-561, 1999.
Nykjaer A, Dragun D, Walther D, Vorum H, Jacobsen C, Herz J, Melsen F, Christensen EI and Willnow TE. An endocytic pathway essential for renal uptake and activation of the steroid 25-(OH) vitamin D3. Cell 96: 507-515, 1999.
Andersen OM, Christensen LL, Christensen PA, Sorensen ES, Jacobsen C, Moestrup SK, Etzerodt M and Thogersen HC. Identification of the minimal functional unit in the low density lipoprotein receptor-related protein for binding the receptor-associated protein (RAP). A conserved acidic residue in the complement-type repeats is important for recognition of RAP. J Biol Chem 2000 Jul 14 ;275 (28 ):21017 -24 275: 21017-21024, 2000.
Andersen OM, Christensen PA, Christensen LL, Jacobsen C, Moestrup SK, Etzerodt M and Thogersen HC. Specific binding of alpha-macroglobulin to complement-type repeat CR4 of the low-density lipoprotein receptor-related protein. Biochemistry 2000 Sep 5 ;39 (35 ):10627 -33 39: 10627-10633, 2000.
Birn H, Fyfe JC, Jacobsen C, Mounier F, Verroust PJ, Orskov H, Willnow TE, Moestrup SK and Christensen EI. Cubilin is an albumin binding protein important for renal tubular albumin reabsorption. J Clin Invest 2000 May ;105 (10 ):1353 -61 105: 1353-1361, 2000.
Chan WL, Shaw PC, Tam SC, Jacobsen C, Gliemann J and Nielsen MS. Trichosanthin interacts with and enters cells via LDL receptor family members. Biochem Biophys Res Commun 2000 Apr 13 ;270 (2 ):453 -7 270: 453-457, 2000.
Graversen JH, Jacobsen C, Sigurskjold BW, Lorentsen RH, Moestrup SK, Thogersen HC and Etzerodt M. Mutational analysis of affinity and selectivity of kringle-tetranectin interaction. GRAFTING NOVEL KRINGLE AFFINITY ONTO THE TETRANECTIN LECTIN SCAFFOLD. J Biol Chem 2000 Dec 1 ;275 (48 ):37390 -6 275: 37390-37396, 2000.
Kristiansen M, Aminoff M, Jacobsen C, de La CA, Krahe R, Verroust PJ and Moestrup SK. Cubilin P1297L mutation associated with hereditary megaloblastic anemia 1 causes impaired recognition of intrinsic factor-vitamin B(12) by cubilin. Blood 2000 Jul 15 ;96 (2 ):405 -9 96: 405-409, 2000.
Sousa MM, Norden AG, Jacobsen C, Willnow TE, Christensen EI, Thakker RV, Verroust PJ, Moestrup SK and Saraiva MJ. Evidence for the role of megalin in renal uptake of transthyretin. J Biol Chem 2000 Dec 8 ;275 (49 ):38176 -81 275: 38176-38181, 2000.
Vorum H, Jacobsen C and Honore B. Calumenin interacts with serum amyloid P component. FEBS Lett 2000 Jan 14 ;465 (2 -3 ):129 -34 465: 129-134, 2000.
Andersen OM, Petersen HH, Jacobsen C, Moestrup SK, Etzerodt M, Andreasen PA and Thogersen HC. Analysis of a two-domain binding site for the urokinase-type plasminogen activator-plasminogen activator inhibitor-1 complex in low-density-lipoprotein-receptor-related protein. Biochemical Journal 357: 289-296, 2001.
Andersen OM, Schwarz FP, Eisenstein E, Jacobsen C, Moestrup SK, Etzerodt M and Thogersen HC. Dominant thermodynamic role of the third independent receptor binding site in the receptor-associated protein RAP. Biochemistry 40: 15408-15417, 2001.
Burmeister R, Boe IM, Nykjaer A, Jacobsen C, Moestrup SK, Verroust P, Christensen EI, Lund J and Willnow TE. A two-receptor pathway for catabolism of Clara cell secretory protein in the kidney. Journal of Biological Chemistry 276: 13295-13301, 2001.
Jacobsen L, Madsen P, Jacobsen C, Nielsen MS, Gliemann J and Petersen CM. Activation and functional characterization of the mosaic receptor SorLA/LR11. Journal of Biological Chemistry 276: 22788-22796, 2001.
Kozyraki R, Fyfe J, Verroust PJ, Jacobsen C, Dautry-Varsat A, Gburek J, Willnow TE, Christensen EI and Moestrup SK. Megalin-dependent cubilin-mediated endocytosis is a major pathway for the apical uptake of transferrin in polarized epithelia. Proceedings of the National Academy of Sciences of the United States of America 98: 12491-12496, 2001.
Kristiansen M, Graversen JH, Jacobsen C, Sonne O, Hoffman HJ, Law SK and Moestrup SK. Identification of the haemoglobin scavenger receptor. Nature 2001 Jan 11 ;409 (6817 ):198 -201 409: 198-201, 2001.
Nykjaer A, Fyfe JC, Kozyraki R, Leheste JR, Jacobsen C, Nielsen MS, Verroust PJ, Aminoff M, de la Chapelle A, Moestrup SK, Ray R, Gliemann J, Willnow TE and Christensen EI. Cubilin dysfunction causes abnormal metabolism of the steroid hormone 25(OH) vitamin D-3. Proceedings of the National Academy of Sciences of the United States of America 98: 13895-13900, 2001.
Birn H, Zhai XY, Holm J, Hansen SI, Jacobsen C, Moestrup SK and Christensen EI. Megalin binds and mediates cellular uptake of the folate binding protein. Journal of the American Society of Nephrology 13: 486A, 2002.
Caterer NR, Graversen JH, Jacobsen C, Moestrup SK, Sigurskjold BW, Etzerodt M and Thogersen HC. Specificity determinants in the interaction of apolipoprotein(a) kringles with tetranectin and LDL. Biol Chem 383: 1743-1750, 2002.
Gburek J, Verroust PJ, Willnow TE, Fyfe JC, Nowacki W, Jacobsen C, Moestrup SK and Christensen EI. Megalin and cubilin are Endocytic receptors involved in renal clearance of hemoglobin. Journal of the American Society of Nephrology 13: 423-430, 2002.
Schmitz C, Hilpert J, Jacobsen C, Boensch C, Christensen EI, Luft FC and Willnow TE. Megalin deficiency offers protection from renal aminoglycoside accumulation. Journal of Biological Chemistry 277: 618-622, 2002.
Conticello SG, Kowalsman ND, Jacobsen C, Yudkovsky G, Sato K, Elazar Z, Petersen CM, Aronheim A and Fainzilber M. The prodomain of a secreted hydrophobic mini-protein facilitates its export from the endoplasmic reticulum by hitchhiking on sorting receptors. J Biol Chem 278: 26311-26314, 2003.
Gburek J, Birn H, Verroust PJ, Goj B, Jacobsen C, Moestrup SK, Willnow TE and Christensen EI. Renal uptake of myoglobin is mediated by the endocytic receptors megalin and cubilin. Am J Physiol Renal Physiol 285: F451-F458, 2003.
Hermey G, Keat SJ, Madsen P, Jacobsen C, Petersen CM and Gliemann J. Characterization of sorCS1, an alternatively spliced receptor with completely different cytoplasmic domains that mediate different trafficking in cells. J Biol Chem 278: 7390-7396, 2003.
Petersen HH, Hilpert J, Militz D, Zandler V, Jacobsen C, Roebroek AJ and Willnow TE. Functional interaction of megalin with the megalinbinding protein (MegBP), a novel tetratrico peptide repeat-containing adaptor molecule. J Cell Sci 116: 453-461, 2003.
Gliemann J, Hermey G, Nykjaer A, Petersen CM, Jacobsen C and Andreasen PA. The mosaic receptor sorLA/LR11 binds components of the plasminogen-activating system and platelet-derived growth factor-BB similarly to LRP1 (low-density lipoprotein receptor-related protein), but mediates slow internalization of bound ligand. Biochem J 381: 203-212, 2004.
Horn IR, Nielsen MJ, Madsen M, Jacobsen C, Graversen JH and Moestrup SK. Generation of a haptoglobin-hemoglobin complex-specific antibody blocking the binding of the complex to CD163 (vol 71, pg 289, 2003). European Journal of Haematology 72: 77, 2004.
Jacobsen C and Moestrup SK. Using Biacore technology to characterize receptors for macromolecular complexes. Biacore Journal 4: 13-15, 2004.
Madsen M, Moller HJ, Nielsen MJ, Jacobsen C, Graversen JH, van den BT and Moestrup SK. Molecular characterization of the haptoglobin.hemoglobin receptor CD163. Ligand binding properties of the scavenger receptor cysteine-rich domain region. J Biol Chem 279: 51561-51567, 2004.
Nykjaer A, Lee R, Teng KK, Jansen P, Madsen P, Nielsen MS, Jacobsen C, Kliemannel M, Schwarz E, Willnow TE, Hempstead BL and Petersen CM. Sortilin is essential for proNGF-induced neuronal cell death. Nature 427: 843-848, 2004.
Petersen HH, Hilpert J, Jacobsen C, Lauwers A, Roebroek AJ and Willnow TE. Low-density lipoprotein receptor-related protein interacts with MafB, a regulator of hindbrain development. FEBS Lett 565: 23-27, 2004.
Petersen SV, Oury TD, Ostergaard L, Valnickova Z, Wegrzyn J, Thogersen IB, Jacobsen C, Bowler RP, Fattman CL, Crapo JD and Enghild JJ. Extracellular superoxide dismutase (EC-SOD) binds to type i collagen and protects against oxidative fragmentation. J Biol Chem 279: 13705-13710, 2004.
Westergaard UB, Sorensen ES, Hermey G, Nielsen MS, Nykjaer A, Kirkegaard K, Jacobsen C, Gliemann J, Madsen P and Petersen CM. Functional organization of the sortilin Vps10p domain. J Biol Chem 279: 50221-50229, 2004.
Birn H, Zhai XY, Holm J, Hansen SI, Jacobsen C, Christensen EI and Moestrup SK. Megalin binds and mediates cellular internalization of folate binding protein. Febs Journal 272: 4423-4430, 2005.
Hvidberg V, Maniecki MB, Jacobsen C, Hojrup P, Moller HJ and Moestrup SK. Identification of the receptor scavenging hemopexin-heme complexes. Blood 2005.
Hvidberg V, Jacobsen C, Strong RK, Cowland JB, Moestrup SK and Borregaard N. The endocytic receptor megalin binds the iron transporting neutrophil-gelatinase-associated lipocalin with high affinity and mediates its cellular uptake. FEBS Lett 579: 773-777, 2005.
Westergaard UB, Kirkegaard K, Sorensen ES, Jacobsen C, Nielsen MS, Petersen CM and Madsen P. SorCS3 does not require propeptide cleavage to bind nerve growth factor. FEBS Lett 579: 1172-1176, 2005.
Sanggaard KW, Sonne-Schmidt CS, Jacobsen C, Thogersen IB, Valnickova Z, Wisniewski HG and Enghild JJ. Evidence for a Two-Step Mechanism Involved in the Formation of Covalent HC.TSG-6 Complexes. Biochemistry 45: 7661-7668, 2006.
Van den Steen PE, Van A, I, Hvidberg V, Piccard H, Fiten P, Jacobsen C, Moestrup SK, Fry S, Royle L, Wormald MR, Wallis R, Rudd PM, Dwek RA and Opdenakker G. The hemopexin and O-glycosylated domains tune gelatinase B/MMP-9 bio-availability via inhibition and binding to cargo receptors. J Biol Chem 2006.
About us
Core facility manager
Core facility technician
At present, the facility offers molecular interaction analysis based on Surface Plasmon Resonance (SPR). In the future, however, we are planning to expand our services to include Isothermal Titration Calorimetry (ITC).