Standing out in the field of IgY Immunotechnology

IgY Purification

 
IgG
IgE
IgY
IgY (ΔFc)
Species

Mammals

Mammals

Birds, Reptiles, Amphibians Lungfish

Anseriform Birds, Amphibians, Lungfish

Molecular Weight (kD)

150

200

180

118

Isoelectric
Point (pI)

6.4-9.0  (Ref. 1)

5.2-5.8  (Ref. 2)

5.7-7.6  (Ref. 3)

5.2-7.3 (Ref. 4)

Extinction
Coefficient*

1.40

1.62  (Ref. 5)

1.36

1.38

No. Constant Domains

4

4

3

2

Carbohydrate
Content (%)

2-4

12

(Ref. 6)

0.6  (Ref. 7)

Hinge Region

Yes

No

No

No

Antigen Valency

2

2

2

2

Major Serum
Antibody

Yes

No

Yes

Yes

Source

Serum

Serum

Serum/EggYolk

Serum/Egg Yolk

 Concentration
(mg/ml)

10-12 

10-4

Chicken:

Serum: 8-10 (Ref. 10)

Yolk: 10-20  (Ref. 9)

Duck:

Yolk: IgY (ΔFc): 3-12

(Ref. 4)

Mammalian Complement binding

Yes

No

No

No

Rheumatoid Factor Binding

Yes

No

No

No

Fc receptor binding

Yes

No

No

No

Mediates
Anaphylaxis

No

Yes

Yes

No

Binding to Protein A

Yes

No

No

No

Binding to Protein G

Yes

No

No

No

Binding to Protein L

Yes

Yes

No  (Ref. 8)

No

* optical density of a 1.0 mg/ml solution at 280 nm.

Other "About IgY" topics:

References:
1. Li G, Stewart R, Conlan B, Gilbert A, Roerth, P, Nair H. (2002) Purification of human immunoglobulin G: a new approach to plasma fractionation. Vox Sanguinis 83 : 332-338.
2. Clerc, M. -F., Granato, D. A.  and M. Horisberger, (1988). Labelling of colloidal gold with IgE. Histochemistry and Cell Biology 89 : 343-349.
3. Dávalos-Pantoja L, Ortega-Vinuesa JL, Bastos-González D, Hidalgo-Alvarez R, (2000)  A comparative study between the adsorption of IgY and IgG on latex particles.  J Biomater Sci Polym Ed. 11:657-73. 
4. Chiou, Victor, 2003. Process for selectively isolating IgY antibodies from egg yolk of an anseriform bird and IgY antibodies obtained thereby. United States Patent Application Publication 20060223986.
5. Liu, F.T., Bohn, J.W., Ferry, E.L., Yamamoto, H., Molinaro, C.A., Sherman, L.A., Klinman, N.R., and Katz, D.H. (1980) Monoclonal dinitrophenyl-specific murine IgE antibody preparation, isolation and characterization. J. Immunol. 124 : 2728-2736.
6. Ohta M, Hamako J, Yamamoto S, Hatta H, Kim M, Yamamoto T, Oka S, Mizuochi T, Matsuura F., 1991, Structures of asparagine-linked oligosaccharides from hen egg-yolk antibody (IgY). Occurrence of unusual glucosylated oligo-mannose type oligosaccharides in a mature glycoprotein.
7. Zimmerman, B., Shalatin, N. and H.M. Grey (1971).  Structural studies on the duck 5.7S and 7.8S immunoglobulins. Biochemistry 10 : 482-488.
8. Nilsson, E. Larsson, A.  (2005) Chicken Anti-Protein L for the Detection of Small Amounts of Protein L in the Presence of IgG.  Hybridoma. 24(2): 112-114
9. Carlander, D. 2002.Avian IgY Antibody. In vitro and in vivo. Acta Universitatis Upsaliensis. Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine 1119.53 pp. Uppsala. ISBN 91-554-5227-2
10. Wang, Y.W., Cherian, G., Sunwoo, H.H and Sim. J.S. 2000. Dietary polyunsaturated fatty acids significantly affect laying hen lymphocyte proliferation and immunoglobulin G concentration in serum and egg yolk. Can. J. Anim. Sci. 80:597-604.

Many researchers are now choosing to make their custom polyclonal antibody in chickens.  There are many reasons to do so. Some choose to immunize a hen and collect the eggs because it is the least invasive way to produce polyclonal antibodies.  The eggs can be stored for up to 3 months in the refrigerator or the yolks separated and frozen in delipidation buffer (Reagent A of the Eggspress IgY Purification Kit).  For some, the attraction is that chickens often produce better antibodies to conserved mammalian proteins.  And for some, choosing hens  means that a life-time supply of polyclonal antibody will be available to them after immunizing a single animal.  One hen can produce as much antibody as 10 – 20 rabbits, and it’s so much easier to collect eggs than bleed 10 rabbits!

Making an IgY antibody in hens isn't that different from producing a rabbit antibody, except that it's easier because bleeding the animal is not necessary.  Perhaps the most difficult part of raising antibodies in chickens is to provide a suitable environment for them.  Hens prefer to roost on sticks at night, lay their eggs in a nesting box and have regular "dust baths" in sand or wood shavings.  At Gallus Immunotech, we provide this environment for our hens.  After all, a happy, unstressed hen is most likely to produce a good IgY antibody!  

Want to try producing your own IgY antibody?  View our chicken immunization protocol here. Want us to make your custom IgY?  Please visit our Custom IgY Production page.

The European Centre for the Validation of Alternative Methods (EVCAM) has reported on a workshop that  validates the benefits of using chickens as laboratory animals to produce polyclonal antibodies.  Click here to view the whole report.

Other "About IgY" topics:

igy-compareIt’s easy to understand why IgY was first categorized as an immunoglobulin G – it’s similar to mammalian IgG (see Figure on right).

Like IgG, IgY also has 2 heavy and 2 light chains, shaped in the characteristic Y shape with an antigen valency of 2.  IgY and IgG share a similar sedimentation coefficient of about 7S and both are eluted from DEAE cellulose with low ionic strength buffers. Functionally, they play a similar biological role as well. Both IgG and IgY are the major immunoglobulins providing defense against infectious agents and appear in blood at high concentrations following synthesis of a higher molecular weight antibody (IgM).

There are striking differences however: There is little or no immunological cross-reactivity between IgY and mammalian IgG.  IgY has a higher molecular weight due to an extra heavy chain constant domain, lacks a well-defined hinge region and has unique oligosaccharide sidechains1.  It has been proposed that the extra domain (CH2) may be the evolutionary precursor to the mammalian IgG hinge region2.  IgY is capable of mediating anaphylactic reactions, a function limited to IgE in mammals.  In fact, other similarities between IgY and IgE (see Comparison of IgG, IgE, IgY and IgY (ΔFc) and Figure to the right), including similar intrachain disulphide bonding in their extra heavy chain domain, have lead some to suggest that IgY is the ancestral molecule to both IgG and IgE. 3  Molecular genetic studies have confirmed this. 4,5,6,7,8

Other "About IgY" topics:

References

1.  Ohta M, Hamako J, Yamamoto S, Hatta H, Kim M, Yamamoto T, Oka S, Mizuochi T., Matsuura F., 1991, Structures of asparagine-linked oligosaccharides from hen egg-yolk antibody (IgY). Occurrence of unusual glucosylated oligo-mannose type oligosaccharides in a mature glycoprotein. Glycoconj J. 8:400-13.
2. Putnam, F.W., Takahashi, N., Tetaert, D., Debuire, B. and Lin, C., 1980, Amino acid sequence of the first constant region of the delta heavy chain of human IgD. PNAS 78 : 6168-6172.
3. Warr, G.W., Magor, K.E. and D.A. Higgins,  (1995) IgY : clues to the origins of modern antibodies. Immunology Today 16 :392–398.
4. Hedges, S.B. (2002) The origin and evolution of model organisms.  Nat. Rev. Genet. 3 : 838-849.
5. Reisz, R.R. and Muller, J. (2004) Molecular timescales and the fossil record: a paleontological perspective. Trends Genet 20 : 237-241.
6. Bininda-Emonds, O.R., Cardillo, M., Jones, K.E., MacPhee, R.D., Beck, R.M., Grenyer, R., Price, S.A., Vos, R.A., Gittleman, J.L. and Purvis, A. (2007) The delayed rise of present-day mammals. Nature 446 : 507-512.
7. Vernersson, M., Aveskogh, M., and Hellman, L. (2004) Cloning of IgE from the echidna (Tachyglossus aculeatus) and a comparative analysis of epsilon chains from all three extant mammalian lineages. Dev. Comp. Immunol. 28 : 61-75.
8. Vernersson, M., Aveskogh, M. Munday, B., and Hellman, L. (2002), Evidence for an early appearance of modern post switch immunoglobulin isotypes in mammalian evolution (II), cloning of IgE, IgG1 and IgG2 from a monotreme, the duckbilled platypus, Ornithorhynchus anatinus. Eur. J. Immunol. 32 : 2145-2155.

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