hen egg yolk antibodies akrum hamdy. in 1893, klemperer first demonstrated that the immunization of...
Post on 20-Dec-2015
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In 1893, Klemperer first demonstrated that the immunization of a hen resulted in the transfer of specific antibodies from the serum to the egg yolk.
For over a hundred years there was no scientific application for this knowledge .
But when the animal welfare became a matter of serious ethical concern for the scientific community, the results of Klemperer have attracted a great attention, particularly since the 1980s.
In the sense of animal welfare, the use of laying hens for antibody production represents a refinement and a reduction in animal use. It is a refinement in that the painful and invasive blood sampling or
scarifying are replaced by collecting eggs .The antibodies productivity in laying hens is nearly 20 times greater than that in rabbits (Schade et al., 2004).
14 HAP Comprise ~96% of the Protein Mass in Plasma
Albumin
IgAIgG total
IgM
Pool Containing Medium- & Low-Abundance Proteins
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TransferrinFibrinogen
2-Macroglobulin
1-Antitrypsin
Haptoglobin
Apolipoproteins A-1 & A-II1-Acid Glycoprotein
SuperMix Approach – Dig Deeper
IgY12 Column
12 HAP-Depleted Fraction (Flow-Through of IgY12 Column: F1)
(Moderate/Low-Abundant Proteins, MAP/LAP)
Protein Mixture as Antigens for
IgY Production
Antigen- Affinity Column
IgY against 12 HAP-Depleted Fraction
(SuperMix)
Eluted Fraction (E1) (12 High-Abundant
Proteins – HAP)
Eluted Fraction (E2) (MAP or Immunoreactive Proteins)
Flow-Through Fraction (F2) (LAP or Non-Immunoreactive Proteins)
SuperMixColumnPlasma Directly Applied to IgY-SuperMix Column
Plasma(P)
IgYs
IgY-Immunoaffinity Separation
1. High avidity of binding
2. Specific removal of the targets
3. Sample size 20-500 l
4. Recyclable >100x
5. Efficacy >95%
6. Minimal loss of non-targets
IgY-BEADS’ FEATURES
Why Chickens & IgYs?
Hen Rabbit
Distance toHuman -205 Myr +72 Myr
Production Accumul Why Chickens & IgYs? ativeInvasive
Monthly 20 Eggs 50 ml Blood
Ab Quantity 100 mg/egg 10 mg/ml
Maintenance Low High
Unit Cost Low High
AffinityMaturation
GeneConversion
Avidity High Good
Somatic Mutation
SepproTip Design
Membrane filter
Stainless steel ring500 µl
SepproTip IgY-12
IgY Microbeads (Seppro® IgY-12)
Membrane filter
SepproTip Process
500 µl Unfractionated Sample (1:33
Dilution)
Binding Washing Eluting Neutralizing Regenerating
Pipetting Up and Down 8 Times Each
Pipetting Up and Down 8 Times Each
Up and Down 10 Times
Blood Samples
Loading 15 µl
Option to Collect the Washes at
Position 2
Collect and Pool Elution Fractions at Positions 3 and 4
Pipetting Up and Down 15 Minutes
Position 1 Wells 1-3 Wells 4-7 Well 8 Wells 9-10
Up and Down 3 Times
Liquid in and stop
SepproTip Content - Customizable
MediaPurposeCapacity
IgY12Partitioning of Top-12 highly-abundant
proteins from human and other primate serum or plasma
15 µl of serum/plasma
IgY-R7Partitioning of Top-7 highly-abundant
proteins from rat serum or plasma10 µl of
serum/plasma
IgY-M7Partitioning of Top-7 highly-abundant
proteins from mouse serum or plasma.10 µl of
serum/plasma
IgY-D11Partitioning of Top-11 highly-abundant
proteins from dog serum or plasma.10 µl of
serum/plasma
SuperEnrich
Enrichment of tissue leakage proteins, cell line-specific
500 ul of plasma/serum
Anti-Rubisco
Removal of Rubisco protein from plant extract (plant biomarker discovery)
0.8 mg of Rubisco
IgY: Less Non-Specific Binding
Mammalian IgG
Chicken IgY
Complement
Rheumatoid Factor
IgM
Fc Receptor
No Binding to Fc Region
Polyclonal Antibody Production
Total IgYs
Bioinformatic Analysis
Expression Vectors
Antigens Ligands
Affinity Purification Quality Control Tests
Chickens and Eggs
Protein Expression Contract Services
•Bioinformatics Analysis
System Selection and Adaptation
Refolding and Solubilization
Protein Expression and Optimization
Scale-Up Production
GLP/cGMP Compliance
Vector Design and Construction
Comparison of IgY and IgG
[35S] LabelingECL Detection
Western Blot Immuno-precipitation
Immuno-staining
IgY IgG
A B C
IgY IgG
Calnuc
Anti-Calnuc IgY(Golgi-Specific)
Body’s defenses against bacteria
Neutralizing antibodies: block viruses from attaching and entering the host
Virus specific antibodies: can act against proteins on the virus surface
Overuse of antibodies can lead to the development of resistant bacteria
conclusion
With the ban on sub-therapeutic antibiotic usage in Europe and the increasingly strictness of the European legislation on food hygiene, passive immunization by oral administration of pathogen-specific hen egg yolk antibody (IgY) may be a
useful and attractive alternative .Furthermore, because of the high yolk antibodies concentration, over 100 mg of antibodies can be obtained from one egg (Akita et al., 1992). Since a laying hen produces approximately 25 eggs per month, 5 g of antibody per month may be obtained from a single laying hen.