Human monoclonal antibody stability and activity at vaginal pH

Philip E. Castle, Daniel A. Karp, Larry Zeitlin, E. Bertrand García-Moreno, Thomas R. Moench, Kevin J. Whaley, Richard A. Cone

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Antibodies can be delivered topically to the vagina to protect against pregnancy and sexually transmitted infections, but the acidity of vaginal secretions (pH 3.5-4.5) might inactivate them. To address this question, both experimental and computational methods were used to evaluate the effects of pH on human monoclonal antibody (MAb) stability and activity. To determine the acid-sensitivity of their antigen binding sites, human MAbs against human sperm (H6-3C4) and gp120 of HIV (1511) were tested by ELISA for binding to human sperm and recombinant gp120, respectively, at pH 3.0-7.0, after storing them for 1 or 20 h at the same pH. Binding was unaltered by acidic pH≥4 even after 20 h, and at pH 3.5 both MAbs retained ≥40% antigen binding activity. A humanized MAb against HSV-2 glycoprotein B expressed both in Chinese hamster ovary (CHO) cells and in soybean cells was incubated for 1 or 24 h at pH 3.5-7.6, brought to neutral pH, and tested for ability to block HSV-2 infection of foreskin fibroblast cells. Loss in blocking activity occurred only when antibodies were incubated at pH 3.5 for 24 h and was independent of the expression cell type. Using empirical structure-based methods, net charge, Z, and electrostatic contributions to free energy, ΔΔGel, were calculated as a function of pH for 1 human and 8 murine F(ab)s. The calculations indicate that Z changes slowly between pH 5.0 and 9.0 and that ΔΔGel is nearly constant between pH 4.0 and 10 for all the F(ab)s and, therefore, human antibodies should remain stable in this pH range. Taken together, our data and empirical calculations suggest that vaginally applied human MAbs are likely to remain stable and active throughout the duration they are likely to reside in the vagina.

Original languageEnglish (US)
Pages (from-to)61-76
Number of pages16
JournalJournal of Reproductive Immunology
Volume56
Issue number1-2
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Monoclonal Antibodies
Vagina
Spermatozoa
Antibodies
Gels
HIV Envelope Protein gp120
Antibodies, Monoclonal, Humanized
Foreskin
Antigens
Human Herpesvirus 2
Sexually Transmitted Diseases
Cricetulus
Static Electricity
Soybeans
Ovary
Fibroblasts
Enzyme-Linked Immunosorbent Assay
Binding Sites
Pregnancy
Acids

Keywords

  • HIV
  • HSV
  • Monoclonal antibodies
  • Plantibodies
  • Sperm
  • Vagina

ASJC Scopus subject areas

  • Immunology
  • Reproductive Medicine

Cite this

Castle, P. E., Karp, D. A., Zeitlin, L., García-Moreno, E. B., Moench, T. R., Whaley, K. J., & Cone, R. A. (2002). Human monoclonal antibody stability and activity at vaginal pH. Journal of Reproductive Immunology, 56(1-2), 61-76. https://doi.org/10.1016/S0165-0378(02)00013-X

Human monoclonal antibody stability and activity at vaginal pH. / Castle, Philip E.; Karp, Daniel A.; Zeitlin, Larry; García-Moreno, E. Bertrand; Moench, Thomas R.; Whaley, Kevin J.; Cone, Richard A.

In: Journal of Reproductive Immunology, Vol. 56, No. 1-2, 2002, p. 61-76.

Research output: Contribution to journalArticle

Castle, PE, Karp, DA, Zeitlin, L, García-Moreno, EB, Moench, TR, Whaley, KJ & Cone, RA 2002, 'Human monoclonal antibody stability and activity at vaginal pH', Journal of Reproductive Immunology, vol. 56, no. 1-2, pp. 61-76. https://doi.org/10.1016/S0165-0378(02)00013-X
Castle, Philip E. ; Karp, Daniel A. ; Zeitlin, Larry ; García-Moreno, E. Bertrand ; Moench, Thomas R. ; Whaley, Kevin J. ; Cone, Richard A. / Human monoclonal antibody stability and activity at vaginal pH. In: Journal of Reproductive Immunology. 2002 ; Vol. 56, No. 1-2. pp. 61-76.
@article{b4504fac5b9644d3831a85951814a7f1,
title = "Human monoclonal antibody stability and activity at vaginal pH",
abstract = "Antibodies can be delivered topically to the vagina to protect against pregnancy and sexually transmitted infections, but the acidity of vaginal secretions (pH 3.5-4.5) might inactivate them. To address this question, both experimental and computational methods were used to evaluate the effects of pH on human monoclonal antibody (MAb) stability and activity. To determine the acid-sensitivity of their antigen binding sites, human MAbs against human sperm (H6-3C4) and gp120 of HIV (1511) were tested by ELISA for binding to human sperm and recombinant gp120, respectively, at pH 3.0-7.0, after storing them for 1 or 20 h at the same pH. Binding was unaltered by acidic pH≥4 even after 20 h, and at pH 3.5 both MAbs retained ≥40{\%} antigen binding activity. A humanized MAb against HSV-2 glycoprotein B expressed both in Chinese hamster ovary (CHO) cells and in soybean cells was incubated for 1 or 24 h at pH 3.5-7.6, brought to neutral pH, and tested for ability to block HSV-2 infection of foreskin fibroblast cells. Loss in blocking activity occurred only when antibodies were incubated at pH 3.5 for 24 h and was independent of the expression cell type. Using empirical structure-based methods, net charge, Z, and electrostatic contributions to free energy, ΔΔGel, were calculated as a function of pH for 1 human and 8 murine F(ab)s. The calculations indicate that Z changes slowly between pH 5.0 and 9.0 and that ΔΔGel is nearly constant between pH 4.0 and 10 for all the F(ab)s and, therefore, human antibodies should remain stable in this pH range. Taken together, our data and empirical calculations suggest that vaginally applied human MAbs are likely to remain stable and active throughout the duration they are likely to reside in the vagina.",
keywords = "HIV, HSV, Monoclonal antibodies, Plantibodies, Sperm, Vagina",
author = "Castle, {Philip E.} and Karp, {Daniel A.} and Larry Zeitlin and Garc{\'i}a-Moreno, {E. Bertrand} and Moench, {Thomas R.} and Whaley, {Kevin J.} and Cone, {Richard A.}",
year = "2002",
doi = "10.1016/S0165-0378(02)00013-X",
language = "English (US)",
volume = "56",
pages = "61--76",
journal = "Journal of Reproductive Immunology",
issn = "0165-0378",
publisher = "Elsevier Ireland Ltd",
number = "1-2",

}

TY - JOUR

T1 - Human monoclonal antibody stability and activity at vaginal pH

AU - Castle, Philip E.

AU - Karp, Daniel A.

AU - Zeitlin, Larry

AU - García-Moreno, E. Bertrand

AU - Moench, Thomas R.

AU - Whaley, Kevin J.

AU - Cone, Richard A.

PY - 2002

Y1 - 2002

N2 - Antibodies can be delivered topically to the vagina to protect against pregnancy and sexually transmitted infections, but the acidity of vaginal secretions (pH 3.5-4.5) might inactivate them. To address this question, both experimental and computational methods were used to evaluate the effects of pH on human monoclonal antibody (MAb) stability and activity. To determine the acid-sensitivity of their antigen binding sites, human MAbs against human sperm (H6-3C4) and gp120 of HIV (1511) were tested by ELISA for binding to human sperm and recombinant gp120, respectively, at pH 3.0-7.0, after storing them for 1 or 20 h at the same pH. Binding was unaltered by acidic pH≥4 even after 20 h, and at pH 3.5 both MAbs retained ≥40% antigen binding activity. A humanized MAb against HSV-2 glycoprotein B expressed both in Chinese hamster ovary (CHO) cells and in soybean cells was incubated for 1 or 24 h at pH 3.5-7.6, brought to neutral pH, and tested for ability to block HSV-2 infection of foreskin fibroblast cells. Loss in blocking activity occurred only when antibodies were incubated at pH 3.5 for 24 h and was independent of the expression cell type. Using empirical structure-based methods, net charge, Z, and electrostatic contributions to free energy, ΔΔGel, were calculated as a function of pH for 1 human and 8 murine F(ab)s. The calculations indicate that Z changes slowly between pH 5.0 and 9.0 and that ΔΔGel is nearly constant between pH 4.0 and 10 for all the F(ab)s and, therefore, human antibodies should remain stable in this pH range. Taken together, our data and empirical calculations suggest that vaginally applied human MAbs are likely to remain stable and active throughout the duration they are likely to reside in the vagina.

AB - Antibodies can be delivered topically to the vagina to protect against pregnancy and sexually transmitted infections, but the acidity of vaginal secretions (pH 3.5-4.5) might inactivate them. To address this question, both experimental and computational methods were used to evaluate the effects of pH on human monoclonal antibody (MAb) stability and activity. To determine the acid-sensitivity of their antigen binding sites, human MAbs against human sperm (H6-3C4) and gp120 of HIV (1511) were tested by ELISA for binding to human sperm and recombinant gp120, respectively, at pH 3.0-7.0, after storing them for 1 or 20 h at the same pH. Binding was unaltered by acidic pH≥4 even after 20 h, and at pH 3.5 both MAbs retained ≥40% antigen binding activity. A humanized MAb against HSV-2 glycoprotein B expressed both in Chinese hamster ovary (CHO) cells and in soybean cells was incubated for 1 or 24 h at pH 3.5-7.6, brought to neutral pH, and tested for ability to block HSV-2 infection of foreskin fibroblast cells. Loss in blocking activity occurred only when antibodies were incubated at pH 3.5 for 24 h and was independent of the expression cell type. Using empirical structure-based methods, net charge, Z, and electrostatic contributions to free energy, ΔΔGel, were calculated as a function of pH for 1 human and 8 murine F(ab)s. The calculations indicate that Z changes slowly between pH 5.0 and 9.0 and that ΔΔGel is nearly constant between pH 4.0 and 10 for all the F(ab)s and, therefore, human antibodies should remain stable in this pH range. Taken together, our data and empirical calculations suggest that vaginally applied human MAbs are likely to remain stable and active throughout the duration they are likely to reside in the vagina.

KW - HIV

KW - HSV

KW - Monoclonal antibodies

KW - Plantibodies

KW - Sperm

KW - Vagina

UR - http://www.scopus.com/inward/record.url?scp=0036304692&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036304692&partnerID=8YFLogxK

U2 - 10.1016/S0165-0378(02)00013-X

DO - 10.1016/S0165-0378(02)00013-X

M3 - Article

VL - 56

SP - 61

EP - 76

JO - Journal of Reproductive Immunology

JF - Journal of Reproductive Immunology

SN - 0165-0378

IS - 1-2

ER -