Skip to main content

Details on Affinity Chromatography- download [pdf] and [ppt]




Affinity chromatography

Affinity chromatography is a method of separating biochemical mixture based on a highly specific interaction between antigen and antibody, enzyme and substrate, receptor and ligand, or protein and nucleic acid.


Back in the year of post-graduation, I have delivered a seminar on Affinity Chromatography.
Its Powerpoint presentation is not so good and I did not include much text in the ppt.
Rather I have decided to include only the informative pictures and I explained the whole topic only with those pictures. (I got good grade for this!)
So, here is this ppt. You can download it if you want!


DOWNLOAD

For preparation of this topic I downloaded A Handbook of Affinity Chromatography.
The principle and methods of this chromatography are well explained in this book.
I am giving link of this book for you. 
Don't forget to download and read! 

DOWNLOAD


Do you know the components of affinity medium?
If not, let's learn about them in short!
Affinity medium is composed of three main components.
1. Matrix:
It is made up of agarose gel.
It serves as an inert matrix to which ligand binds directly or indirectly.

2. Ligand:
Ligand is a component which binds to biomolecules (enzymes, proteins, etc).
This component is specific for each kind of biomolecule.

3. Spacer arm:
Binding site is located deep within a molecule. 
Spacer arm helps the molecule to bind with ligand.
When binding, length of the spacer arm is an important thing.


Why the length of spacer arm is important?

Importance of length of spacer arm
Importance of length of spacerarm | Source - Wikimedia Commons

The image is self-explanatory, isn't it?
In short, if length of spacer arm is very short, then molecule will not bind properly with ligand.
Ligand will not reach at the binding site of molecule.

If spacer arm is very long, non-specific binding may occur.

For this reason the length of spacer arm should be optimum.




Labels:

Comments

Post a Comment

Popular posts from this blog

Aromatic waters & Tinctures: Monophasic liquid dosage forms

AROMATIC WATERS Aromatic waters are the saturated solutions of volatile oils or other aromatic substances.   Methods of preparation of aromatic waters: Distillation: Crude drug/material is placed in sufficient amount of purified water in flask. After heating water forms a steam which is condensed to obtain a condensate which contain aromatic principles. This method is expensive, tedious and time consuming. Examples: Orange flower water NF, Strong Rose water NF. Solution method: In this method, the volatile oil is shaken for 15 min with sufficient quantity (500 times) of water to make a solution. The resultant solution is kept aside for 12 hrs then filtered through wet filter paper.  Examples: Dill water, Peppermint water, Chloroform water, Camphor water, etc. Alternate solution method: The volatile oil is mixed with an inert adsorptive material ( Talc, Kieselghur, purified siliceous earth) then 1 ltr of purified water is added and agitated for 10 min. Resulting solution is...
Post a Comment
Read more

Linctuses: Monophasic Liquid Dosage Form

  LINCTUSES What is a Linctus? Linctuses are sweet, viscous liquid oral preparations containing medicinal substances which have demulscent, expectorant or sedative properties. These contain high proportions of syrup and glycerin which exert demulscent effect on the mucous membrane of throat. In order to obtain prolonged local action, linctuses should be administered slowly in undiluted form. Formulation of Linctus  1. Vehicle : Syrups are mostly used as vehicles in the linctuses. Tolu syrup is preferred in cough preparations due to its aromatic odor and flavour. Glycerol syrup and invert syrup are also used as vehicles. Linctus preparation for diabetic patients contain sorbitol syrup (vehicle). Medicament is first dissolved in little amount of water and then added into vehicle. Syrups are viscous and contain less amount of water, this limits (affects) the dissolution of medicament in syrup. 2. Additives: Colouring agents : Amaranth, Erythrosin, Tartrazine. Flavouring agents : ...
Post a Comment
Read more

UV-Visible spectroscopy: Basics

Ultraviolet Spectroscopy In this anaytical technique, ultraviolet radiations are used. Wavelength ranges of radiations: Visible light : 400 - 800 nm Near UV radiations : 200 - 400 nm Far/ Vacuum UV : below 200 nm Generally, we carry out UV analysis in near UV region of radiations. PRINCIPLE : Electronic transitions When a molecule absorbs UV radiations, the electronic excitation occur where electrons go from lower to higher energy state.  Electronic excitation causes the electron to go from electron bonding orbital to antibonding orbital. After some time electron returns to its original state (stable). While returning to its stable form (into bonding orbital) it releases excess energy. That energy is measured by the detector to access the wavelength (λmax) /absorption pattern. TRANSITIONS: σ → σ* (126-135 nm) n → σ* (180-200 nm) Ï€ → Ï€* n → Ï€* ALLOWED TRANSITION:  Ï€ → Ï€* FORBIDDEN TRANSITIONS:    n → Ï€* ENERGY OF VARIO...
Post a Comment
Read more