OFERTA DE EMPLEO “ANALISTA CIENTÍFICO EN BIOFÍSICA”

AFFINImeter es una spin-off de la Universidad de Santiago de Compostela que se dedica a desarrollo herramientas de análisis basadas en técnicas calorimétricas para su aplicación en las primeras fases del desarrollo de fármacos. Actualmente AFFINImeter ofrece software y servicios a empresas como Pfizer, Novartis, Lilly o Moderna, así como a muchas de las universidades más prestigiosas del mundo. Además, AFFINImeter es socio de consorcio MOSBRI (MOlecular-Scale Biophysics Research Infrastructure) financiado por la Unión Europea para promocionar el acceso a técnicas biofísicas a la comunidad científica europea.

RESPONSABILIDADES:

  • Realización de experimentos de Calorimetría de Titulación Isotérmica (ITC) y otras técnicas biofísicas (Uv-vis, Fluorescence, NMR, etc…)
  • Apoyo en el diseño y validación de una base de datos biofísicos.
  • Asesoramiento científico a clientes internacionales y prospección comercial.

QUÉ OFERTAMOS:

  • Contrato indefinido a tiempo completo.
  • Incorporación inmediata.
  • Salario bruto ~22K -26K€
  • Posibilidad de realizar tu doctorado industrial.
  • Posibilidad de teletrabajo

QUÉ NECESITAMOS:

  • Licenciados/as en Física, Química, Bioquímica o Biotecnología.
  • Nivel alto de inglés, oral y escrito.
  • Iniciativa y autonomía en el trabajo.
  • Se valorarán conocimientos de programación y diseño de base de datos.
  • Disponibilidad para viajar.

Envía tu Curriculum y carta de presentación a info@affinimeter.com

Download AFFINImeter ITC Advanced for Windows

We’ve released AFFINImeter ITC Advanced 1.2.0 for Windows

Do you want to install and use AFFINImeter in your computer? Now you can.

Go ahead and download AFFINImeter ITC for Windows right now (only for 64 bits computers).
Upon download, all users will get a FREE 30 days trial version for AFFINImeter ITC Advanced, including: KinITC, ITC and TA data analysis, Model builder, Design complex fitting and simulation projects for ITC, and a new tool: the rawdata experiments simulator.

Users who purchased a license for the web version have the same license for the Windows version. Just contact us at info@affinimeter.com to activate it.

 

Release notes for 1.2.0 (Update is recommended):

Important updates have been performed in the windows version of the AFFINImeter-ITC software. 

The analysis has implemented the accurate determination of concentrations recently published by Philippe Dumas: (see https://www.biorxiv.org/content/10.1101/512780v1.full).

This update breaks backwards compatibility: this means that all previously created projects (.affprojects) will not be compatible with this version (they will not load in the application or will throw some errors). The previously processed itc/ta raw data should be processed again to update the accurate calculation of concentrations in the equilibrium isotherm.

Other problems/issues fixed:

  • [Fixed] Fitting Projects Output may show incorrect results while performing complex fittings.
  • [Added] Fitting Projects now have an option to download a ZIP file containing all charts in the project, and also some new and specific ones.
  • [Added] Presentation of local minima for fitting projects was improved.
  • [Added/Fixed] Y labels, ticks and names
    • Y values/ticks will have an exponential format instead of the SI prefixes
    • Some Y label names were fixed
  • [Fixed] “Auto” setting in KinITC will stay active after deleting points in the Isotherm, if user the user has it active. Previously, after changing/deleting points in the isotherm, that setting was lost.
  • [Added] Included more references in the Help Section

AFFINImeter Biophysical Services



“All models are wrong but some are useful”

George E. P. Box (1919-2013) British Mathematician

 

Seeking the structure and energetics of the molecules is bearing relevance the drug discovery field.
The biophysical techniques such us ITCNMRMST… are used to unravel many stages of the drug discovery process. Specifically, those are important in the early stages of the drug discovery where a thousand of compounds that may be potential candidates as a medical treatment, are tested.

Our aim is to help scientists whose research is relevant to the drug discovery field and bring to them our expertise on molecular interaction applying biophysical techniques. We truly believe that we make the difference in your research. For this reason, we want to present our services:

 

 

 

Binding Characterization

We use biophysical and computation methods for elucidating complex binding mechanisms.

 

 

 

 

Data Analysis

We have developed our exclusive tools for thermodynamic, kinetic and structural characterization of drug-target interactions.

 

 

 

 

 

Software Development​
We design ad-hoc analysis software to improve your workflow.

 

 

 


 

Visit our web

KinITC: Transforming the ITC roles in Drug Discovery.

Since the kinetic information is the more relevant thing to look at in drug discovery, a typical question comes to our minds:

Which technique should we choose to get kinetic information?

This question doesn´t have an easy answer because many factors should be taken into account to make this decision, but lately, one technique is rising up: Isothermal titration calorimetry and more specifically, the KinITC method developed by P Duma et al [1].

 

In AFFINImeter we have implemented the KinITC method that is making possible the measurement of binding kinetics, kon and koff rate constants, through the analysis of the thermogram (s) obtained from a traditional ITC experiment (s). (more…)

Open position: Finance Controller

We are looking for an experienced and well-rounded Finance Controller to be part of an energetic team dedicated to improving our highly innovative scientific software and services. This role will be based in Santiago de Compostela and it is an excellent opportunity for a passionate professional willing to reach the next level in his/her career.

You will report directly to the CEO and support him in Treasury and Cash Management, Cash balance forecasting and develop business plans & financial projections. Analysis and support preparation of all recurring finance activities, providing financial analysis to support decision making.

The main responsibility of the Finance Controller is to develop and manage strategies, operations and policies related to the treasury function, including cash management, borrowing, and payment processing. This role will be also responsible to establish relationships with banks, payment gateways and other external funding sources.

Main Responsibilities:

  • Finding public and private sources of funding.
  • Develop business plans including financial projections. Coordinate and direct the preparation of the budget and financial forecasts on a quarterly and annual basis.
  • Participation in the design and implementation of a KPI information system for all areas of the company, operational audits, cost/benefit analysis, information services, document management systems and procedures, reporting and other systems management functions.
  • Develop, improve and issue monthly financial records.
  • Be the point of contact for any accounting/control related issues and queries.
  • Manage the financial close process.
  • Act as a key resource provider to the auditors for annual audit and for all the non-trading companies and other financial institutions
  • Improve financial status by analysing results; monitoring variances; identifying trends; recommending actions to management.
  • Other duties as necessary International start-up specialised in scientific software.

Background and Skills

  • Bachelor’s degree in Accounting or Finance. CPA/ACCA/MBA or equivalent preferred.
  • 8+ years of experience, 3+ in a senior position in an analysis or senior financial accountant’s level, preferably in software companies and/or startup environments.
  • Independent worker and analytical thinker with the ability to conduct research, data analysis and resolve complex problems.
  • Strong organizational skills and attention to detail.
  • A “hand’s on” business partner attitude is required; who enjoys challenges and thrives in a demanding environment. Ability to handle multiple tasks concurrently.
  • PC skills with Microsoft products with advanced Excel capabilities.
  • Excellent written and verbal communication skills.
  • Fluent English level.
  • Openwork environment with young, dynamic and outstanding international teams.

We offer:

  • Salary: based on interview, bonuses.
  • Real ownership and results.
  • And a competitive, friendly, international team!

 

Please contact staff@affinimeter.com

 

Analysis of Fluorescence Polarization competition assays with AFFINImeter

Fluorescence Polarization competition assays are widely used in numerous research fields to determine the affinity of unlabeled ligands that compete with a fluorescent probe for binding with the same macromolecule. Herein, we show how to perform a thoughtful analysis of these experiments with AFFINImeter for spectroscopic techniques, to elucidate, not just IC50 values, but also a quantitative direct measurement (KA) of the binding affinity.

 

Introduction


Fluorescence polarization (FP) is a powerful technique that nowadays is widely utilized in high-throughput screening (HTS) and drug discovery (1). In FP assays monitoring a binding event is possible mainly because this technique is sensitive to changes in molecular weight. The assay requires of a fluorescent molecule (the probe) that is excited by plane-polarized light. For small molecules, the initial polarization decreases rapidly due to rotational diffusion during the lifetime of the fluorescence, which results in low fluorescence polarization. When the small fluorescent molecule binds to a larger molecule (and consequently of slower rotation) an increased fluorescent polarization is observed (Fig. 1).
Advances in experimental aspects of the technique like assay design and fluorescent probes is enabling the application of FP to increasingly complex biological processes (1).

Fig.1. The principle of fluorescence polarization (2).

 

Frequently, a competition displacement assay format is used in FP experiments where a fluorescent labelled molecule bound to a macromolecule is displaced by an unlabeled molecule with the consequent decrease of polarization. This assay yields a quantitative measure of IC50 values (half maximal inhibitory concentration) of the unlabeled competitors. However, IC50 does not provide a direct measure of affinity and the calculation of the binding constant (KA) is often desirable. In this sense, the software AFFINImeter Spectroscopy offers exclusive analysis tools to perform a thoughtful analysis of FP competition assays, to directly deliver values of binding constants of the interaction between the competitor and the macromolecule. As an illustrative example, we present herein the use of AFFINImeter Spectroscopy for the analysis of FP competition assays to characterize oligosaccharide –protein interactions.

 

FP competition assays to study the binding between midkine and chondroitin sulfate-like tetrasaccharides


Midkine is a cytokine which biological activity is regulated by its binding with glycosaminoglycans (GAGs), such as heparin and chondroitin sulfate. To better understand these recognition processes Pedro M. Nieto et al. have recently reported the binding of a series of synthetic chondroitin sulfate-like tetrasaccharides with midkine, using FP competition assays (3). First, the direct binding of midkine and a fluorescein labelled heparin hexasaccharide (fluorescent probe) was monitored in a direct FP titration (Fig. 2a). Second, FP competition assays were performed, in which the polarization of samples containing fixed concentrations of midkine and fluorescent probe was recorded in the presence of increasing concentrations of different synthetic sugars to obtain the corresponding IC50 values (Fig. 2b) (3).

 

 

 

 

 

 

 

Fig.2. a) Binding curve of the titration of fluorescent probe with midkine 3a;b) Representative competition curve (semi-log plot) of an FP competition assay where the fluorescent probe is displaced from a pre-formed complex with midkine by the competitor.

 

Analysis of FP competition assays with AFFINImeter Spectroscopy


The determination of KA of the interaction of the unlabeled ligand with the macromolecule in an FP competition assay is possible if the corresponding curve is analyzed using a competitive binding model where two equilibria are described, one between the free species and the macromolecule–probe complex and one between the free species and the macromolecule–competitor complex. In this analysis, the KA value of the interaction between probe and macromolecule can be fixed (as it can be previously calculated from the direct binding experiment) in order to determine KA of the competitor-macromolecule complex. Yet, a more robust approach consists of a global analysis of direct and competitive curves where constraints between experimental parameters are imposed. Using this approach, we have used AFFINImeter Spectroscopy in the analysis of data from the direct measurement of the fluorescent labelled hexasaccharide binding to midkine and together with data from the displacement assay using an unlabeled, synthetic disaccharide as a competitor (4).

An AFFINImeter fitting project was generated where the curve from the direct titration and two curves from replicates of the competitive assay were included. A 1:1 simple binding model and a competitive model were used to fit direct and competitive data, respectively (Fig. 3).

Fig.3. Schematic representation of the binding models used to fit the FP data.5 These models can be easily built with the “model builder” tool of AFFINImeter where a) in the 1:1 model “M” represents the species sensitive to binding (fluorescent probe) and “A” represents the titrant (midkine); b) in the competitive model “M” is the sensitive species (probe), A is the titrant (competitor) and B is a third species involved in the event (midkine in a preformed complex with the probe).

 

In the analysis, the fitting parameters considered were the KA of each complex, the signal value of the unbound state (s0), and the maximum signal change (Δsmax) of the midkine-probe complex formation. Δsmax of the midkine-competitor complex is equal to zero because the competitor is not fluorescent. Since midkine-probe binding is present in both models, restrictions were made considering that KA and Δsmax of the 1:1 model (FS↔MA) are equal to the same parameters describing this equilibrium in the competitive model (FS↔MB). Besides, s0 was common between replicates of the competitive assay. It is worth to mention that the curves analyzed are the mean of three replicate experiments and the corresponding standard deviation of the curve data points are also considered in the fitting process.  The global analysis performed in this way returned the KA of the interaction of midkine with the probe, (3.09 ± 0.18)*107 M-1, and with the competitor, (5.30 ± 0.52)*106 M-1 (Fig. 4). Additionally, AFFINImeter automatically generates the species distribution plot, valuable in the interpretation of results. The species distribution plot of Fig. 4c shows the displacement of the probe by the unlabeled disaccharide in the competition assay.

 

Fig.4. a) Global analysis of FP curves from the direct titration of probe (10 nM) with midkine (12 – 750 nM) and from a competitive assay consisting of a sample with probe (10 nM) and midkine (63 nM) where the competitor is added at increasing concentrations (0 – 20 mM). Curves from two replicates of the competitive assay were used in the analysis. All the polarization values are the average of three replicate wells and the error bars represent the corresponding standard deviation; b) values of KA, s0 and Δsmax determined for each equilibrium; c) species distribution semi-log plot of the competitive assay.

 

Conclusions


This case study exemplifies the utility of AFFINImeter Spectroscopy in the analysis of FP competitive binding assays. The advantages of using this software rely on the possibility to obtain reliable KA values of the binding between competitor and macromolecule from a global analysis where KA and Δsmax of the probe-macromolecule complex, are shared parameters between curves (they are not pre-determined fixed parameters). Besides, standard deviation between replicates is taken into account in the fitting process. Ultimately, these tools provide a more robust analysis and reliable characterization of binding interactions monitored through competitive assays.

 

Try AFFINImeter Spectroscopy

Acknowledgements


We would like to thank Dr Pedro Nieto Mesa and Dr José Luis de Paz Carrera from the Institute of Chemical Research (IIQ) of the Spanish National Research Council (CSIC), for kindly share with us the FP data presented herein.

 

References & Notes


1 Hall, M.; Yasgar, A.; Peryea, T.; Braisted, J.; Jadhav, A.; Simeonov, A.; Coussens, N. Fluorescence Polarization Assays In High-Throughput Screening And Drug Discovery: A Review. Methods and Applications in Fluorescence 2016, 4, 022001.

2 This figure has been taken from http://glycoforum.gr.jp/science/word/glycotechnology/GT-C06E.html.

3 a) Solera, C.; Macchione, G.; Maza, S.; Kayser, M.; Corzana, F.; de Paz, J.; Nieto, P. Chondroitin Sulfate Tetrasaccharides: Synthesis, Three-Dimensional Structure And Interaction With Midkine. Chemistry – A European Journal 2016, 22, 2356-2369; b) Maza, S.; Gandia-Aguado, N.; de Paz, J.; Nieto, P. Fluorous-Tag Assisted Synthesis Of A Glycosaminoglycan Mimetic Tetrasaccharide As A High-Affinity FGF-2 And Midkine Ligand. Bioorganic & Medicinal Chemistry 2018, 26, 1076-1085.

4 The competitor is a synthetic disulfated disaccharide described in ref. 3b as compound 18.

5 These model were created with the “model builder” tool, exclusive of AFFINImeter. For more information go to AFFINImeter knowledge center.

Working with AFFINImeter models based on a stoichiometric equilibria approach

The appropriate design and use of binding models in AFFINImeter pass through an understanding of the nomenclature that the software uses to describe a given experimental setup and the species that take part in the assay.
AFFINImeter contemplates the presence of up to three species participating in the experiment: 1) the titrant, or compound placed in the syringe; 2) the titrate, or compound in the calorimetric cell and 3) a co-solute or third compound that can be in the syringe and/or in the cell. These species are labelled in the reaction builder as follows:

As illustrated in Fig. 1, M always refers to the compound placed in the calorimetric cell and A always refers to the compound in the syringe. B always refers to a third component that can be in the syringe (Fig. 1b), in the cell (Fig. 1c) or in both places at once (Fig. 1d):

 

Once the nomenclature of  AFFINImeter is clear, you can download this note that our scientific team have prepared to understand how our users can be working with models based on a stoichiometric equilibria approach:

Download

You can also check our latest post where we describe “The concepts of stoichiometric and site binding constants”

 

AFFINImeter is on sales!

 

This month we have prepared a special offer just to let you exploit the full potential of AFFINImeter:

 15 % off in our Advanced plans*.

Select the technique AFFINImeter Spectroscopy and/or AFFINImeter ITC.

 

          

 

… Use this coupon in our online shop and get right away your discount!

ER7DS30

To use our online shop it will be necessary to have an AFFINImeter account, to create one click here.

AFFINImeter Shop

Or if you prefer we can send you an official quotation, click on the button below:​

Get a Quotation

 

 

*The promotion is available until May 31st and applicable for all AFFINImeter Advanced plan (ITC and/or Spectroscopy) which duration is equal or greater 1 year. The advanced account includes Global fitting, access to complex binding models, Kinetic information (KinITC tool available for AFFINImeter ITC), orthogonal approach (Multitechnique analysis available in AFFINImeter Spectroscopy)...

 

 

 

AFFINImeter new version!

Last week we announced a release of our new software: AFFINImeter for Spectroscopic Techniques.  Together with this release, we have also improved several aspects of the AFFINImeter software, see what is new in our new version:

1. As we have already mentioned, in this version included a new technique: Spectroscopy (and all its variants: UV-vis, InfraRed, Fluorescence, Circular Dichroism… among others)

  • Just upload new data, select Spectroscopy technique and follow the wizard to import your data.
  • Data must be uploaded in CSV format.
  • Create projects of Fitting and Global fitting with any model you want.

2. Recent activity shows you the latest modified/created files or projects of any technique

 

3. Now AFFINImeter will use the maximum width of your screen to show your data/projects, for a better experience using wide-screen monitors.

4. Navigation path visible in all views

Fixed problems:

1. Fixed recursively deleting a folder and its contents

2. More validations while selecting a model for NMR technique (and Spectroscopy also):

  • In NMR and SPEC, you cannot use a model that only contains A in its reactions. Now more here

3. Multiple minor usability improvements and performance tweaks.

4. Multiple bug fixes and issues suggested by our users have been implemented and resolved.

 

What does it look like to you?

It might look like a plain hat or an elephant being eaten by a boa constrictor, but actually, the shape of this curve looks like a typical spectrum of UV-Vis determined by changes in the intensity of this energy and also it represents the logo of our new software:

AFFINImeter for spectroscopic techniques!

 

Hence, we want to take this opportunity to launch AFFINImeter Spectroscopy software.

It is a new software for the processing and advanced analysis of binding isotherms obtained from different spectroscopic techniques: 1D-NMR, Uv-vis, fluorescence, circular dichroism, differential scanning fluorimetry…

 

…and from now on, you will be able to start using it:

Get a free trial


To get started, we have prepared some scientific material as an example of how to use AFFINImeter Spectroscopy:


We have prepared a special offer to celebrate the launch of AFFINImeter for Spectroscopic techniques.
Using this code coupon you will get a 25% off*, to buy through our online shop:

Spectroscopy_coupon


 
 
 You can also ask for a quotation here 
 
 
*The discount will be applied until 2018/03/03.