Bioconjugation technologies and services

Transcription

1 Bioconjugation technologies and services (0)

2 Contents Antibody and protein labeling - Lightning-Link Oligonucleotide labeling kits - Thunder-Link PLUS Gold nanoparticle conjugation kits - InnovaCoat GOLD Colloidal gold Pre-conjugated gold nanoparticles LATEX conjugation kits ATPase & GTPase assay kits Phopshate detection reagents 18 B.I.T.S. Immunoassay kits Antibody conjugates Purification kits and accessory products Custom services

3 About us Innova Biosciences are world leading experts in easy-to-use, cuttingedge bioconjugation technologies and services. We have the flexibility to support scientists from academia through to commercial manufacturing, developing and supplying reagent products to make science easier, both off-the shelf and to customer specification. Our products are based on our antibody and protein labeling, nanoparticle, oligonucleotide labeling, and phosphate detection enzyme activity assay technologies. All of our product ranges are of the highest quality and are designed to streamline R&D and manufacturing processes, whilst also reducing overall project costs by cutting costs associated with e.g. material waste, in-house equipment, and staff time, providing increased return on investment compared to traditional methods. Our ever expanding portfolio, including the Lightning-Link, InnovaCoat and Thunder-Link brands, is available to research laboratories, pharmaceutical, biotechnology and diagnostic companies worldwide. USA and Canada UK and rest of world +44 (0) (0)

4 Antibody and protein labeling Lightning-Link Lightning-Link is the world s fastest, easiest to use and most efficient antibody conjugation method, offering antibody and protein labeling with only 30 seconds hands-on time and no separation steps. 30 seconds hands-on time 100% antibody recovery Over 40 available labels using same chemistry Antibody & protein labeling Fully scalable from µg to grams The Lightning-Link conjugation process We ve been really impressed with Innova s products. Their biotin kit is simple to use and proved more durable than many competitors kits, and we ve seen significant improvements in reproducibility since we started to use them regularly. We re looking forward to continuing to work with the Lightning Link technology. Dr Jake Micallef, CSO of VolitionRx 4

5 Fluorescent Label Lightning-Link product code Maximal Absorbance (nm) Excitation colour Suggested Maximal Excitation Laser Emission (nm) Line (nm) AMCA N/A DyLight N/A Atto DyLight PerCP ** PerCP/Cy ** DyLight Fluorescein ** R-Phycoerythrin ** 498, 544, , 532, PE/Texas Red ** 498, 544, , 532, PE/Atto ** 498, 544, , 532, PE/Cy ** 498, 544, , 532, PE/Cy ** 498, 544, , 532, PE/Cy ** 498, 544, , 532, Atto B-Phycoerythrin ** Cyanine Dye Rhodamine DyLight Atto FluoProbes * 630 DyLight * 629 Texas Red * 616 DyLight , 635, Atto , 635, FluoProbes647H , 635, Cyanine Dye , 635, Allophycocyanin ** , 635, APC/Cy ** , 635, APC/Cy ** , 635, DyLight , 635, Cyanine Dye * 705 DyLight * 716 Atto * 724 DyLight * 794 DyLight * 798 Emission Color (R-)PE has three maxima, and all can be used. The optimal will depend on the application. * This Laser Line is some distance from the Maximal Absorbance, so performance will be compromised if this dye is used with the suggested Laser Line. ** These kits require a 3 hour incubation time (instead of 15 minutes). Antibody & protein labeling Enzyme labeling kits Product code Biotin/Streptavidin labeling kits Product code Lightning-Link Alkaline Phosphatase (AP) Lightning-Link Glucose Oxidase Lightning-Link Horseradish Peroxidase (HRP) Lightning-Link Biotin A, Biotin B* , Lightning-Link Rapid Biotin A, Biotin B* , Lightning-Link Avidin Lightning-Link Streptavidin * Biotin A - intended for assays in which a streptavidin-labeled detection reagent will be used. Biotin B - optimised for assays in which the biotinylated protein is captured by streptavidin immobilized on a surface (0)

6 ELISA data Direct vs indirect labeling Alkaline Phosphatase anti-human IgG (Fc) Conjugates analysed by ELISA Antibody & protein labeling Anti-human IgG monoclonal antibody was purchased pre-conjugated to alkaline phosphatase from a commercial supplier. The same antibody was purchased in un-conjugated form and then conjugated to alkaline phosphatase using a Lightning-Link kit. The Lightning-Link conjugate demonstrates enhanced titre and sensitivity. Immunohistochemistry data Dual staining using Lightning-Link conjugates Dual staining of CD3 and CD20 with antibodies directly conjugated using Lightning-Link kits. Mouse anti-human CD3 was conjugated with alkaline phosphatase, and mouse anti-human CD20 was conjugated using HRP. Both antibodies were used simultaneously to stain formalinfixed, paraffin embedded human lymph node. CD20 positive cells are visualised using DAB (Brown) and CD3 cells with Fast Red (Red). Direct labeling of monoclonal antibodies enables straightforward dual-color staining in IHC, which is otherwise a highly complex and time-consuming technique. 6

7 Flow cytometry data Lightning-Link vs ready conjugated antibody Lightning-Link RPA-T4 clone for human CD4, conjugated to RPE Ready conjugated antibody RPA-T4 clone for human CD4, conjugated to RPE RPA-T4 clone for human CD4, conjugated to FITC RPA-T4 clone for human CD4, conjugated to FITC Mouse monoclonal antibodies specific for human cell surface markers were purchased from a commercial source in both unconjugated and ready conjugated formats. The unconjugated antibodies were linked to R-PE or FITC using a Lightning-Link kit, and the conjugates were compared in flow cytometry staining of human peripheral blood lymphocytes. The blue curve shows unstained cells. Western blot data Direct vs indirect labeling Antibody & protein labeling Primary antibody directly conjugated to HRP using Lightning-Link shows enhanced sensitivity in western blotting compared to the traditional indirect technique (0)

8 Oligonucleotide Labeling Kits Thunder-Link PLUS The Thunder-Link PLUS oligonucleotide conjugation kit enables the simple and rapid conjugation of antibodies to oligonucleotides. The kit is quick and easy to use, overcoming time consuming and lengthy protocols associated with standard conjugation methods. Any oligo sequence may be used Fast oligo conjugation - only 1 hour! High antibody and oligo recovery Oligonucleotide conjugation Robust and flexible clean-up procedure Works with antibody fragments and other proteins ns Products Product code 3 reaction + control (a single reaction to conjugate 100µg antibody) reaction + control (each reaction to conjugate 100µg antibody)

9 The Thunder-Link PLUS conjugation process Custom services Oligonucleotide conjugation From finding the best performing conjugate for your specific assay through to generating your optimised conjugates at your required scale and ratio, our experienced conjugation scientists can help. See page 26 for more information on our custom antibody- oligo microoptimisation and custom conjugation services (0)

10 Gold Nanoparticle Conjugation Kits InnovaCoat GOLD InnovaCoat GOLD is a one-step gold nanoparticle conjugation kit for antibodies, proteins and peptides or any other biomolecule with an amine group. It allows easy conjugation of gold nanoparticles for use in R&D applications and for the development and manufacturing of diagnostic kits. Proprietary surface coat Ultra-stable Potential enhanced assay sensitivity Gold nanoparticle conjugation Covalently link antibodies and analytes Different surface chemistries The InnovaCoat GOLD process Having made our own passively adsorbed antibody-coated gold colloid for some years then more recently moving to a high OD, no-spin method, we were a little reluctant to try out another method quite so soon. However the extreme simplicity and rapidity of the InnovaCoat GOLD covalent conjugation, combined with the excellent functionality and consistency of the final product greatly helped us make the change. We now use less colloid per test in a demanding, room temperature stored, liquid formulation and are finding the development of new rapid tests to be significantly easier. Phil Goodwin, UK Biotechnology Professional 10

11 InnovaCoat GOLD has a unique surface coat that completely shields the metallic core of the nanoparticle from the external environment, eliminating colloidal instability. 2nm InnovaCoat surface 40nm particle Size Product Common Applications 10nm 20nm 40nm 60nm 80nm Target Group InnovaCoat GOLD Amine InnovaCoat GOLD Maleimide Thiol InnovaCoat GOLD Hydrazide Aldehyde Transmission EM Scanning EM Lateral Flow Assay FRET Western Blot Dot Blot Light Microscopy Dynamic Force Microscopy DLS Flow Cytometry Gold nanoparticle conjugation (0)

12 Lateral flow data InnovaCoat GOLD vs traditional techniques Gold nanoparticle conjugation Antibody conjugation using InnovaCoat GOLD vs traditional passive conjugation techniques with uncoated gold nanoparticles, showing both enhanced signal intensity and improved specificity. 40nm gold particles were labeled with anti-iga antibody, and used to measure IgA concentration in a lateral flow inhibition assay, with IgA bound to a lateral flow strip. The graph shows a comparison of test and control line signal intensities between biotin conjugated gold nanoparticles from Competitor 1 or 2 and covalently conjugated InnovaCoat GOLD-Biotin on lateral flow test strips. 12

13 InnovaCoat GOLD alternative chemistries As the InnovaCoat surface is firmly anchored to the gold nanoparticle and is resistant to extreme conditions, it is therefore very easy to introduce derivatives of the main technology. Standard Labeling InnovaCoat GOLD InnovaCoat GOLD Hydrazide InnovaCoat GOLD Maleimide Optimization services & kits We can quickly develop multiple antibody conjugates using a range of proprietary methods. InnovaCoat GOLD kits are also available if you wish to carry out your own rapid screening of panels of antibodies. We can further optimize conjugates using an extended collection of non-catalogue materials and other chemistries if required, and carry out preliminary testing of conjugates. Custom coating services We appreciate that you may like to benefit from a simplified production process using a one-step covalent approach but still use your own gold nanoparticles. We have tested a broad range of gold nanoparticles and have successfully applied the InnovaCoat surface in every case. Gold nanoparticle conjugation (0)

14 Colloidal gold Ultra high quality gold nanoparticles Our gold nanoparticles have been rigorously tested against market-leading brands and shown to be superior. These gold nanoparticles boast many benefits including: Uniform spherical shape Narrow size distribution Ultra high quality Batch-to-batch consistency Available at high concentrations to simplify assay development Available in bulk quantities Colloidal gold Our ultra-high quality colloidal gold is currently available in 10nm and 40nm sizes in the concentrations shown below. 10nm 40nm Products Concentration 1OD 10OD 15OD 20OD 10nm 40nm 20ml 100ml 10ml 100ml 14

15 Pre-conjugated gold nanoparticles InnovaCoat GOLD Conjugates Our pre-conjugated gold nanoparticles are manufactured using our InnovaCoat GOLD nanoparticles which have a proprietary surface coat that covalently bind the detection protein providing highly stable conjugates. Products 10nm 20nm 40nm 80nm Target group InnovaCoat GOLD Streptavidin Biotin InnovaCoat GOLD Biotin Streptavidin InnovaCoat GOLD Goat Anti-Mouse Mouse IgG InnovaCoat GOLD Goat Anti-Rabbit Rabbit IgG InnovaCoat GOLD Protein A IgG InnovaCoat GOLD Protein G IgG Gold nanoparticle conjugates For large quantities of InnovaCoat GOLD conjugates or colloidal gold please contact us for pricing: e: t: +44 (0) (0)

16 Latex bead conjugation LATEX conjugation kits Our LATEX bead conjugation kit is a one-step kit for covalently conjugating antibodies, proteins and peptides (or any other biomolecule with an amine group) to specially treated latex beads without the need for extensive optimisation. Quick and easy to use - conjugates ready to use in 35 minutes Our latex is specially treated and resistant to aggregation No extensive ph optimisations required! The LATEX conjugation kit process 16

17 Lateral flow data Figure 1. Lateral Flow Assay of our specially-treated latex beads conjugated to a monoclonal anti- CRP antibody (mab 1) titrated against CRP. 2 ng/ml CRP can easily be detected. Figure 2. Lateral Flow Assay of our specially-treated latex conjugated to the monoclonal anti-crp antibody (mab 2) titrated against CRP in buffer or CRP spiked CRPdepleted serum (100 % serum). The latex behaves similarly in serum and buffer, and no aggregation or non-specific binding is seen. Figure 3. Multiplexed Latex. The three colors of latex each conjugated to a different antibody or protein and forming a line either by a direct binding event (red and black latex) or a sandwich assay binding event (blue latex). The three colors of latex demonstrate no aggregation or background staining. Latex Bead Conjugation Products 1 reaction Midi kit 4 reaction Mini kit 10 reaction Mini kit LATEX Conjugation Kit - Blue 400nm LATEX Conjugation Kit - Red 400nm LATEX Conjugation Kit - Black 400nm (0)

18 ATPase and GTPase assay kits Phosphate detection reagents At the heart of this range of assay kits is PiColorLock TM, a universal phosphate detection reagent for all phosphate-generating enzymes which forms colored complexes that are stable for hours. ATPase & GTPase assay kits Colorimetric (non-radioactive) assay Special additives ensure complete suppression of non-enzymatic backgrounds with acid-labile substrates(e.g. ATP, GTP) Ultra stable phosphate-dye complexes which last for hours Compatible with almost any assay buffer Compatible with multiple enzyme targets in drug discovery Long shelf life Competitor assays suffer from several problems including reagent precipitation. PiColorLock ensures high stability of the colored dye-phosphate complexes (green color). 18

19 PiColorLock Competitor C PiColorLock Competitor A Competitor B PiColorLock has been designed to have a large linear range, thus reducing the need for sample dilution. Competitors products are linear over a much narrower range of concentrations. ATP has been incubated in three phosphate detection reagents. A steadily rising non-enzymatic background signal is seen with competitor reagents, whereas PiColorLock gives baseline readings. Products PiColorLock detection reagent Product: PiColorLock is a phosphate detection reagent for measuring phosphatases, ATPases, GTPases and other enzymes that release inorganic phosphate (Pi). The reagent comes with with an Accelerator, Stabiliser and Pi Standard ideal for high throughput screening. ATPase and GTPase Assay kits Product: , These non-radioactive colorimetric assay kits use a 96 well format. All the necessary reagents are supplied for measuring enzyme activity. 2 and 5 plate kits available. ATPase & GTPase assay kits Lyophilized ATP & GTP vials Product: ATP GTP Ultra high quality ATP and GTP to ensure the lowest possible assay background. Just reconstitute by adding water, avoid multiple freezethaw cycles to ensure high performance ATP and GTP in your assay. PiBind resin Product: PiBind resin provides a remarkably quick and easy way to remove contaminating Pi from buffers therefore reducing high assay backgrounds. The resin works over a broad range of ph values and is unaffected by many commonly used buffer additives (0)

20 B.I.T.S. Immunoassay Kits Developed by Mologic in partnership with Innova Biosciences We have teamed up with Mologic, a specialist diagnostic development company, to offer a range of high-quality immunoassay kits. The B.I.T.S. ELISA and lateral flow kits are easy to use and provide fast, accurate, and high quality immunoassays which require no sample preparation. They contain everything required for ELISA and lateral flow assays, including high quality, stable enzyme or gold conjugates, produced using our world-leading conjugation technology. Immunoassay kits Reliable, rigorously validated assays Competitively priced without compromising on quality Batch-to-batch reproducibility Speed of assays allows you time to do more research Simple procedure minimises operator error 20

21 The B.I.T.S. assay range currently focuses on inflammatory biomarkers. The first products launched are the Desmosine ELISA Kit, fmlp ELISA Kit, and the Human A1AT Lateral Flow Kit, with more kits due for release soon. Human A1AT Lateral Flow kit* The B.I.T.S. TM Lateral Flow kit for Human Alpha-1 antitrypsin (A1AT) provides fast, accurate assays of A1AT in plasma and urine. fmlp ELISA kit* The B.I.T.S. TM fmlp ELISA kit provides fast, accurate results for levels of fmlp in human urine, with a 1.5 hour assay time. Desmosine ELISA kit* The unique B.I.T.S. TM desmosine ELISA kit has been developed to provide assays of desmosine in urine. *Kits are for research use only Coming soon... ELISA Kits PLA2G1B A1AT (Alpha-1-antitrypsin) HNE (Human neutrophil elastase) Adiponectin Leptin CRP Procalcitonin MMP8 TNFa Fibrinogen TIMP1 TIMP2 IL6 Lateral Flow Kits TIMP1 TIMP2 Cystatin C Adiponectin Leptin Immunoassay kits (0)

22 Antibody conjugates Antibody conjugates The availability of antibodies directly conjugated to the enzyme or fluorescent dye of choice remains limited in many areas. We have developed our antibody conjugation technology to offer a range of biomarker-specific antibodies conjugated to any one of our 20 different labels, each generated using our well established Lightning-Link technology to ensure optimum conjugation performance. All of our antibody conjugates are mouse anti-human, are stored at 4 C, shipped at room temperature and have a shelf life of 12 months (6 months for tandem labels). Visit our website or ask us for our antibody conjugate catalogue to view our extensive library of cardiac biomarkers. 22

23 The full list of specificities covered by the Innova cardiac marker panel is shown below. In each case multiple antibody clones are available please enquire for further details. Antibody Adiponectin Albumin BNP NT-proBNP Calcitonin CD40 ligand (CD40L) CRP Cystatin-C D dimer Fatty acid binding protein GPBB IGFBP-4 KIM Lectin-like oxidized LDL receptor 1 (LOX-1) Myeloperoxidase (MPO) Myoglobin PAPP-A dimer PAPP-A Procalcitonin Retinol Binding Protein 4 S100 beta Serum amyloid A Troponin C Troponin Complex Troponin I - dephosphorylated Troponin I - phosphorylated Troponin I Troponin T Label Unconjugated InnovaCoat GOLD HRP Alkaline Phosphatase Glucose Oxidase Biotin Atto 390 Fluorescein Atto488 PerCP PerCP/Cy5.5 RPE RPE-Cy5 RPE-Cy7 Rhodamine Cy3 Atto565 Cy5 Texas Red Atto633 APC APC/Cy5.5 APC/Cy7 Atto700 Antibody conjugates (0)

24 Accessory Products Purification kits and affinity resins Commercially available antibodies often contain substances (e.g. BSA, glycine, tris, and azide) that interfere with labeling reactions. We have developed a range of purification kits which allow for easy and rapid purification of antibodies from any buffer formulation for use with our Lightning-Link, InnovaCoat and LATEX conjugation kits. The range also includes products for antibody purification from tissue culture supernatant (TCS), serum and ascites fluid; for concentration of antibodies and proteins; and for performing buffer exchanges. Accessory products Purified antibody compatible with our conjugation kits (no dialysis required) Multiple species including mouse and rat specific kits High recovery ( 80-90%) Easy-to-use All components included Scale-up available Our range of purification kits can be used to: 99 Concentrate antibodies and other proteins 99 Exchange your buffer (ph, salt concentration) 99 Remove non-protein additives including:»» BSA»» Sodium Azide»» Gelatin»» Glycine»» Tris 24

25 Maleimide Activated Labels, Thiolation Kits For those who have molecules with thiol groups, our ultra-stable freezedried maleimide activated labels are designed to facilitate the production of conjugates. To complement this range, our Thiolation Kits allow thiols to be introduced easily into proteins and other biomolecules. Our stabilised thiol detection reagent allows the successful introduction of thiols (or presence of thiols) to be confirmed and quantified prior to conjugation with a maleimide activated label. Immunogen Kits Our Imm-Link TM immunogen kits allow the conjugation of your hapten to BSA, KLH or Ovalbumin. Simply add a solution of the hapten to a lyophilised mixure containing the carrier protein and the required conjugation chemistry. There are 3 different chemistries available - carboxyl, sulfhydryl and amine - perfect for preparing antigens for immunization. Accessory products Secondary reagents Our high-quality, stable streptavidin, protein A and biotin conjugates complement our range of labeling technologies and provide researchers with even more flexibility and choice of detection methods for their immunoassay. Our streptavidin and protein A conjugates are produced using our unique Lightning-Link bioconjugation technology, therefore providing high quality, stable conjugates for use in your application (0)

26 Custom Services Custom services Micro-optimisation services Find the best performing conjugate for your specific assay! Optimising your antibody conjugates can be time consuming and difficult and can require significant amounts of antibody and label. Our antibody and oligo micro-optimisation services provide a fast and efficient way to optimise your antibody or oligo conjugates using very small quantities of your antibody. A choice of 3 different scales of conjugate 50ug, 100ug and 500ug Over 30 different labels available Antibody clean-up prior to conjugation Evidence of successful conjugation provided Scalable Fast turnaround time Full technical support Conjugation services Our conjugation service is performed by in-house conjugation experts, providing you with the required antibody conjugate or antibody-oligo conjugate at your agreed conjugation ratio and scale. What s included? Generation of conjugate at required scale Antibody clean-up prior to conjugation (removal of BSA or azide) Antibody buffer exchanged in to a conjugation friendly buffer if required Evidence of successful conjugation provided 26

27 Bulk orders Innova Biosciences can supply almost all its catalogue products in quantities that will fit your own research and manufacturing needs. Our bulk products have the same high level of performance and quality as our standard catalogue products only on a larger scale. Bulk orders can be provided in a format that best suits your needs. Custom Pack Sizes We are able to offer our customers the luxury of choosing their own pack size. Our highly skilled manufacturing team can provide you with your very own bespoke pack size. For example we can offer our standard Lightning-Link conjugation kits including unique formulations in almost any size you may require up to 100mg, ideal for those customers who need to perform large scale conjugations on a regular basis. Inventory Control We can offer a batch validation and reservation service. Although all our products are manufactured to the highest possible standards and quality checked to stringent parameters, there are still customers that wish to avoid potential lot to lot variation. For these customers we offer our inventory control service. With this service we give you the opportunity to test different lots of our product and the option to reserve a specific batch of your choice. Benefits of these services include: Quantities and pricing to suit your requirements Packaging that best suits your needs Fast turnaround time High level of quality and performance Reproducibility Guaranteed supply Full technical support If you would like any more information about this service, or have any technical questions please contact us at technical.enquiries@innovabiosciences.com. Custom services (0)

28 Innova Biosciences Ltd. Babraham Hall Babraham Cambridge CB22 3AT United Kingdom US Toll Free: t: +44 (0) e: w: Innova Biosciences products are sold for research purposes only, and our terms and conditions of sale include a limited use license to our Intellectual Property for internal research applications. Commercial use, such as use within manufacturing, re-sale to third parties, or incorporation into kits, requires a separate written agreement, conferring relevant additional rights, with Innova Biosciences. Lightning-Link, InnovaCoat and Thunder-Link are registered trademarks of Innova Biosciences Cy and CyDye are trademarks of GE Healthcare DyLight is a registered trademark of Thermo Fisher Scientific Inc. and its subsidiaries FluoProbes is a registered trademark of interchim Texas Red is a registered trademark of Life Technologies Corporation