Effect of phosphorylation of ovalbumin on adsorption by aluminum-containing adjuvants and elution upon exposure to interstitial fluid.
The phosphate content of commercial ovalbumin was increased from 1.8 to 3.2 mol PO(4)/mol ovalbumin by conjugation of phosphoserine and reduced to 1.2 or 0.14 mol PO(4)/mol ovalbumin by treatment with potato acid phosphatase. The four ovalbumin samples were completely adsorbed by aluminum hydroxide adjuvant due to electrostatic attraction of the negatively charged ovalbumin and the positively charged aluminum hydroxide adjuvant as well as by ligand exchange of phosphate groups with surface hydroxyl groups. Elution from aluminum hydroxide adjuvant upon exposure to interstitial fluid was inversely related to the degree of phosphorylation of the ovalbumin. The ovalbumin sample containing 3.2 mol PO(4)/mol ovalbumin did not elute while the ovalbumin sample containing 0.14 mol PO(4)/mol ovalbumin eluted completely from aluminum hydroxide adjuvant during exposure to interstitial fluid for 30 min. Adsorption of the four ovalbumin samples by aluminum phosphate adjuvant was directly related to the degree of phosphorylation of ovalbumin. Adsorption was due to ligand exchange as an electrostatic repulsive force operated between the negatively charged ovalbumin samples and the negatively charged aluminum phosphate adjuvant. The potential for ligand exchange decreased as the phosphorylation of ovalbumin decreased. Elution upon exposure to interstitial fluid was inversely related to the degree of phosphorylation and was more extensive than observed for aluminum hydroxide adjuvant. Adsorption of ovalbumin by aluminum-containing adjuvants and elution upon exposure to interstitial fluid can be controlled by the degree of phosphorylation of both ovalbumin and the aluminum-containing adjuvant.
Morefield
Garry L
GL
Department of Industrial and Physical Pharmacy, Purdue University, 575 Stadium Mall Drive, Room 124, West Lafayette, IN 47907-2091, USA.
Jiang
Dongping
D
Romero-Mendez
Ilia Z
IZ
Geahlen
Robert L
RL
Hogenesch
Harm
H
Hem
Stanley L
SL
D000135:Acid Phosphatase / Q000378:metabolism; D000276:Adjuvants, Immunologic / Q000737:chemistry* / Q000493:pharmacokinetics; D000327:Adsorption; D000536:Aluminum Hydroxide / Q000737:chemistry* / Q000493:pharmacokinetics; D000818:Animals; D045604:Extracellular Fluid / Q000378:metabolism*; D066298:In Vitro Techniques; D010047:Ovalbumin / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D010768:Phosphoserine / Q000737:chemistry; D012756:Sheep
D000276:Adjuvants, Immunologic; D010768:Phosphoserine; D000536:Aluminum Hydroxide; D010047:Ovalbumin; D000135:Acid Phosphatase