Reduction of endotoxin contamination of various crude vaccine materials by gram-negative bacteria using aminated poly(gamma-methyl L-glutamate) spherical particles.
We describe a method for the removal of endotoxins from various crude antigen solutions originating from gram-negative bacteria using aminated poly(gamma-methyl L-glutamate) (PMLG) spherical particles. The aminated PMLG adsorbents showed high affinity for various purified endotoxins at an ionic strength of mu = 0.1. The endotoxin-adsorbing capacity of the adsorbent increased with increase in the amino-group content of the adsorbent. The adsorbent (3.2 meq/g amino-group content) showed the highest affinity for endotoxin at ionic strengths ranging from mu = 0.025-0.8. The adsorption of Bordetella pertussis antigen to the adsorbent decreased with increasing amino-group content of the adsorbent at an ionic strength of mu = 0.2. The adsorption of B. bronchiseptica protein to the adsorbent increased with increasing amino-group content of the adsorbent, but decreased with increasing ionic strength. The adsorbent (3.2 meq/g of amino-group content) selectively reduced endotoxin in crude antigen solutions originating from gram-negative bacteria, B. pertussis, B. bronchiseptica and Pasteurella multocida, even at a high ionic strength (mu = 0.2-0.4) without affecting the recovery of the protective antigens.
Sakata
M
M
Department of Applied Chemistry, Faculty of Engineering, Kumamoto University, Japan.
Kawai
T
T
Ohkuma
K
K
Ihara
H
H
Hirayama
C
C
D000327:Adsorption; D000586:Amination; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D001428:Bacterial Vaccines / Q000737:chemistry / Q000302:isolation & purification*; D016950:Bordetella bronchiseptica / Q000276:immunology; D001886:Bordetella pertussis / Q000276:immunology; D004340:Drug Contamination; D004731:Endotoxins / Q000737:chemistry* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006090:Gram-Negative Bacteria / Q000276:immunology*; D006168:Guinea Pigs; D008863:Microspheres; D009994:Osmolar Concentration; D011099:Polyglutamic Acid / Q000031:analogs & derivatives* / Q000737:chemistry
D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D004731:Endotoxins; C030315:poly-gamma-methylglutamate; D011099:Polyglutamic Acid