In-vitro Techniques in Biochemistry and Pharmacology

Course content

The following practical elements are included: primary cell cultures, neurotransmitter transporters, receptor activation (GPRC and ligand-gated ion-channel receptors), receptor binding, electrophysiology, second messenger measurements, FRET, enzyme fractionation, enzyme inhibition, smooth muscle contraction, CytP450 and drug metabolism, separation of lipophilic drug metabolites, use of radioisotopes, drug effects on isolated mitochondria.

Education

MSc Programme in Pharmacy or Pharmaceutical Sciences (Danish programmes cand.pharm and cand.scient.pharm) - elective

MSc Programme in Medicinal Chemistry - elective

MSc Programme in Pharmaceutical Sciences (English programme)- elective 

MSc Programme in Environmental Science (SCIENCE) - restricted elective

 

Learning outcome

By the end of the course, the students will have acquired the competences to:

Knowledge

  • demonstrate a firm knowledge of in vitro pharmacology 
  • understand common in vitro experimental methodologies as applied in pharmacology
  • demonstrate a basic knowledge of pharmacodynamic theory
  • demonstrate a basic understanding of radioisotope theory in scintillation counting

 

Skills

  • The students will acquire practical and theoretical abilities and skills in pharmacological in vitro research techniques that are applicable in both academic and biotech/pharmaceutical drug discovery research environments. The specific aim is to acquire the abilities and practical skills required to be able to work in a pre-clinical drug discovery research environment and/or communicate proficiently with other professionals.
  • prepare cultures of primary mouse neurons
  • execute neurotransmitter uptake experiments employing cultured neurons
  • measure cAMP (second-messenger) in cultured cells by FRET
  • employ isolated mouse vas deferens to measure the effect of morphine-like pain-relieving drugs
  • employ two-electrode voltage clamp (TEVC) electrophysiology technique as a functional assay of drugs affecting the operation of ion channels
  • employ radioligand binding assays in drug investigations and interpret the resulting pharmacologial data
  • purify an enzyme employing affinity chromatography
  • perform enzyme inhibition assays
  • analyze cytochrome P450-mediated metabolism of a radiolabeled drug in a liver homogenate employing thin layer chromatography (TLC) and subsequent scintillation counting
  • isolate crude mitochondria from a mouse liver homogenate
  • employ isolated mitochondria to perform a simple in vitro screen of toxicity related to mitochondrial function of unknown drugs.

 

Competences

  • work with and/or communicate proficiently with other professionals within the field of in vitro pharmacology
  • design simple in vitro assays of drug activity including choosing the proper model system and assay conditions
  • calculate and interpret pharmacological data (dose-response relationships)
  • apply knowledge of in vitro pharmacology to evaluate pharmacological data in relation to the drug discovery process

Lab exercises: 44 hours
Tutorials: 11 hours

  • Lab manual (Pickering and Hansen)
  • Receptor theory notes (Sheykhzade and Pickering)
  • Radioisotope theory notes (Hansen and Pickering)

Students should have completed bachelor’s level lecture course(s) in biochemistry and/or pharmacology as well as a lab course in chemistry and/or biochemistry.

The schedule for this course is not compatible with that of Clinical Pharmacy Practice (SFKKB9011U).
This course also runs in week 42.
Attendance in all lab exercises and tutorials is mandatory.

ECTS
7,5 ECTS
Type of assessment
Written examination, 2 hours under invigilation
Examination design
The course examination will consist of 20 short-answer questions covering the practical and theoretical aspects of all the laboratory exercises. All written answers will be given an equal weighting and then the course leaders will assess the overall examination performance level.
Aid
Without aids

There is access to the following at the exam on Peter Bangs Vej:

  • Office (Word, Excel, Onenote and Powerpoint)
  • IO2 – the digital pen
  • Panoramic Viewer
  • Paint
  • Calculator – Windows' own
  • R – Statistical programme
  • ITX MC – multiple choice programme
  • Adobe reader

 

usb is not allowed

Marking scale
7-point grading scale
Censorship form
No external censorship
Criteria for exam assessment

To achieve the grade 12 the student must be able to:

Knowledge

  • demonstrate a firm knowledge of in vitro pharmacology 
  • understand common in vitro experimental methodologies as applied in pharmacology
  • demonstrate a basic knowledge of pharmacodynamic theory
  • demonstrate a basic understanding of radioisotope theory in scintillation counting

 

Skills

  • prepare cultures of primary mouse neurons
  • execute neurotransmitter uptake experiments employing cultured neurons
  • measure cAMP (second-messenger) in cultured cells by FRET
  • employ isolated mouse vas deferens to measure the effect of morphine-like pain-relieving drugs
  • employ two-electrode voltage clamp (TEVC) electrophysiology technique as a functional assay of drugs affecting the operation of ion channels
  • employ radioligand binding assays in drug investigations and interpret the resulting pharmacologial data
  • purify an enzyme employing affinity chromatography
  • perform enzyme inhibition assays
  • analyze cytochrome P450-mediated metabolism of a radiolabeled drug in a liver homogenate employing thin layer chromatography (TLC) and subsequent scintillation counting
  • isolate crude mitochondria from a mouse liver homogenate
  • employ isolated mitochondria to perform a simple in vitro screen of toxicity related to mitochondrial function of unknown drugs.

 

Competences

  • work with and/or communicate proficiently with other professionals within the field of in vitro pharmacology
  • design simple in vitro assays of drug activity including choosing the proper model system and assay conditions
  • calculate and interpret pharmacological data (dose-response relationships)
  • apply knowledge of in vitro pharmacology to evaluate pharmacological data in relation to the drug discovery process

Single subject courses (day)

  • Category
  • Hours
  • Practical exercises
  • 44
  • Colloquia
  • 22
  • Lectures
  • 2
  • Preparation
  • 88
  • Exam Preparation
  • 50
  • English
  • 206